• BMB Reports
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• v.56 no.3
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• pp.172-177
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• 2023

BEST family is a class of Ca²⁺-activated Cl- channels evolutionary well conserved from bacteria to human. The human BEST paralogs (BEST1-BEST4) share significant amino acid sequence homology in the N-terminal region, which forms the transmembrane helicases and contains the direct calcium-binding site, Ca²⁺-clasp. But the cytosolic C-terminal region is less conserved in the paralogs. Interestingly, this domain-specific sequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in the BEST channels is still poorly understood. Thus, we aimed to understand the functional role of the C-terminal region in the human and mouse BEST1 channels by using electrophysiological recordings. We found that the calcium-dependent activation of BEST1 channels can be modulated by the C-terminal region. The C-terminal deletion hBEST1 reduced the Ca²⁺-dependent current activation and the hBEST1-mBEST1 chimera showed a significantly reduced calcium sensitivity to hBEST1 in the HEK293 cells. And the C-terminal domain could regulate cellular expression and plasma membrane targeting of BEST1 channels. Our results can provide a basis for understanding the C-terminal roles in the structure-function of BEST family proteins.

• Environmental Engineering Research
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• v.21 no.4
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• pp.341-349
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• 2016

To improve the utilization rate of construction waste as mine backfilling materials, this paper investigated the feasibility of using recycled powder as mine paste backfilling cementitious material, and studied the pozzolanic activity of recycled construction waste powder. In this study, alkali-calcium-sulfur served as the activation principle and an orthogonal test plan was performed to analyze the impact of the early strength agent, quick lime, and gypsum on the pozzolanic activity of the recycled powder. Our results indicated that in descending order, early strength agent > quick lime > gypsum affected the strength of the backfilling paste with recycled powder as a cementitious material during early phases. The strength during late phases was affected by, in descending order, quick lime > gypsum > early strength agent. Using setting time and early compressive strength as an analysis index as well as an extreme difference analysis, it was found that the optimal ratio of recycled powder cementitious material for mine paste backfilling was recycled powder:quick lime:gypsum:early strength agent at 78%:10%:8%:4%. X-ray diffraction analysis and scanning electron microscope were used to show that the hydration products of recycled powder cementitious material at the initial stages were mainly CH and ettringite. As hydration time increased, more and more recycled powder was activated. It mainly became calcium silicate hydrate, calcium aluminate hydrate, etc. In summary, recycled powder exhibited potential pozzolanic activities. When activated, it could replace cementitious materials to be used in mine backfill.

• Molecular Medicine Reports
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• v.20 no.3
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• pp.2476-2483
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• 2019

Atopic dermatitis (AD ) is an inflammatory skin disorder caused by immunological dysregulation and genetic factors. Whether the expression levels of cytokine and skin barrier protein were altered by S100 calcium binding protein A8 (S100A8) and S100A9 in human keratinocytic HaCaT cells was examined in the present study. Alterations of cytokine expression were examined by ELI SA following treatment with S100A8/9 and various signal protein-specific inhibitors. Activation of the mitogen activated protein kinase (MAPK) pathway and nuclear factor (NF)-κB was evaluated by using western blotting and an NF-κB activity test, respectively. The expression levels of interleukin (IL )-6, IL- 8 and monocyte chemoattractant protein-1 increased following treatment with S100A8 and S100A9, and the increase was significantly blocked by specific signaling pathway inhibitors, including toll-like receptor 4 inhibitor (TLR 4i), rottlerin, PD98059, SB203580 and BAY-11-7085. Extracellular signal-regulated kinase (ER K) and p38 MAPK pathways were activated in a time-dependent manner following treatment with S100A8 and S100A9. Phosphorylation of ER K and p38 MAPK were blocked by TLR 4i and rottlerin. S100A8 and S100A9 induced translocation of NF-κB in a time-dependent manner, and the activation of NF-κB was inhibited by TLR 4i, rottlerin, PD98059 and SB203580. In addition, S100A8 and S100A9 decreased the expression of skin barrier proteins, filaggrin and loricrin. These results may help to elucidate the pathogenic mechanisms of AD and develop clinical strategies for controlling AD.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.241-249
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• 2017

Plasma membrane hyperpolarization associated with activation of $Ca^{2+}$-activated $K^+$ channels plays an important role in sperm capacitation during fertilization. Although Slo3 (slowpoke homologue 3), together with the auxiliary ${\gamma}^2$-subunit, LRRC52 (leucine-rich-repeat-containing 52), is known to mediate the pH-sensitive, sperm-specific $K^+$ current KSper in mice, the molecular identity of this channel in human sperm remains controversial. In this study, we tested the classical $BK_{Ca}$ activators, NS1619 and LDD175, on human Slo3, heterologously expressed in HEK293 cells together with its functional interacting ${\gamma}^2$ subunit, hLRRC52. As previously reported, Slo3 $K^+$ current was unaffected by iberiotoxin or 4-aminopyridine, but was inhibited by ~50% by 20 mM TEA. Extracellular alkalinization potentiated hSlo3 $K^+$ current, and internal alkalinization and $Ca^{2+}$ elevation induced a leftward shift its activation voltage. NS1619, which acts intracellularly to modulate hSlo1 gating, attenuated hSlo3 $K^+$ currents, whereas LDD175 increased this current and induced membrane potential hyperpolarization. LDD175-induced potentiation was not associated with a change in the half-activation voltage at different intracellular pHs (pH 7.3 and pH 8.0) in the absence of intracellular $Ca^{2+}$. In contrast, elevation of intracellular $Ca^{2+}$ dramatically enhanced the LDD175-induced leftward shift in the half-activation potential of hSlo3. Therefore, the mechanism of action does not involve pH-dependent modulation of hSlo3 gating; instead, LDD175 may modulate $Ca^{2+}$-dependent activation of hSlo3. Thus, LDD175 potentially activates native KSper and may induce membrane hyperpolarization-associated hyperactivation in human sperm.

• Advances in materials Research
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• v.4 no.3
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• pp.133-144
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• 2015

The aim of this investigation is to prepare geopolymer resin by alkali activation of ceramic waste (AACW) with different sodium hydroxide (NaOH) and liquid sodium silicate (LSS) concentrations. In order to prepare geopolymer cement, AACW was replaced by 10 and 30 % by weight (wt.,) of concrete waste (CoW) as well as 10 and 30 wt., % ground granulated blast-furnace slag (GGBFS). The results showed that, the compressive strength of AACW increases with the increase of activator content up to 15:15 wt., % NaOH: LSS. All AACW hardened specimens activated by 3:3 (MC6), 6:6 (MC12), 12:12 (MC24) and 15:15 wt., % (MC30) NaOH: LSS destroyed when cured in water for 24h. The MC18 mix showed higher resistivity to water curing. The results also showed that, the replacement of AACW containing 9:9 wt., % NaOH: LSS (MC18) by 10 (MCCo10) and 30 (MCCo30) wt., % CoWdecreased the compressive strength at all ages of curing. In contrast, the MCCo10 mix showed the lower chemically combined water content compared to MC18 mix. The MCCo30 mix showed the higher chemically combined water content compared to MC18 and MCCo10 mixes. The compressive strength and chemically combined water of all AACWmixes containing GGBFS (MCS10 and MCS30) were higher than those of AACWwith no GGBFS (MC18). As the amount of GGBFS content increases the chemically combined water increases. The x-ray diffraction (XRD) proved that as the amount of CoWcontent increases, the degree of crystallinity increases. Conversely, the replacement of AACW by GGBFS leads to increase the amorphiticity character. The infrared spectroscopy (FTIR) confirms the higher reactivity of GGBFS compared to CoW as a result of successive hydration products formation, enhancing the compaction of microstructure as observed in scanning electron microscopy (SEM).

• Microbiology and Biotechnology Letters
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• v.14 no.4
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• pp.285-291
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• 1986

Immobilized Candida lipolytica cells were prepared by entrapping the whole cells in calcium alginate gel. To enhance citric acid productivity, immobilized cells were Incubated with activation medium in fluidized-bed reactors. When the activation was done in batch operation, maximum citric acid productivity appeared in a much shorter time than in continuous operation. Activated immobilized cells were enhanced about 10-fold in citric acid production relative to non-activated immobilized cells. The productivity of citric acid was also influenced by bead size. When Immobilized cells were reacted in a fluidized-bed reactor with the same quantity of cells, the citric acid productivity was increased as the bead size was decreased.

• Molecules and Cells
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• v.44 no.2
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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.

• The Korean Journal of Physiology
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• v.27 no.2
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• pp.175-183
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• 1993

There are many report suggesting that influx and intracellular calcium concentration $([Ca^{2+}]_i)$ are related to cell signalling in various cells. However, it has not been reported that calcium channel activation is affected by the substances involved in signal transduction pathways in the mouse eggs. In this study, the effects of isoprenaline (ISP) and cyclic AMP on calcium influx through calcium channels were investigated to show their relationship with the signal transduction process in unfertilized mouse eggs. Using whole cell voltage clamp techniques, calcium currents, elicited by the depolarizing pulses of 300 ms duration (from -50 mV to 50 mV in 10 mV increments) from a holding potential of -80 mV, were recorded. The current-voltage (I-V) relation of calcium currents was shown to be bell-shaped; the current began to activate at -50 mV and reached its maximum $(-1.33{\pm}0.16\;nA:\;mean{\pm}S.E.,\;n=7)$ at -10 mV, then decayed at around 50 mV. Calcium currents were fully activated within $7\;ms{\sim}20\;ms$ and completely inactivated 200 ms after onset of the step pulse. ISP within the concentration ranges of $10^{-8}\;M{\sim}10^{-4}\;M$ dose-dependently increased the amplitude calcium current. The permeable cyclic AMP analogue,8-bromocyclic AMP, also increased its maximal amplitude by 46ft at $10^{-5}\;M$, while protein kinase inhibitor (PKI), which is known to inhibit 0.02 phosphorylating units of cyclic AMP-dependent protein kinase (PKA) per microgram decreased calcium currents. Currents recorded in the presence of PKI were resistant to increase by the application of $10^{-5}\;M$. Also, PKI inhibited the calcium current increase elicited by ISP treatment. These results suggest that $\beta-adrenergic$ regulation of the calcium channel is mediated by the cAMP-dependent protein kinase. This signal transduction pathway might play a role in regulating $[Ca^{2+}]_i$, level due to the increase of calcium influx in mouse eggs.

• Biomolecules & Therapeutics
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• v.26 no.5
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• pp.439-445
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• 2018

T-type calcium channels are low voltage-activated calcium channels that evoke small and transient calcium currents. Recently, T-type calcium channels have been implicated in neurodevelopmental disorders such as autism spectrum disorder and neural tube defects. However, their function during embryonic development is largely unknown. Here, we investigated the function and expression of T-type calcium channels in embryonic neural progenitor cells (NPCs). First, we compared the expression of T-type calcium channel subtypes (CaV3.1, 3.2, and 3.3) in NPCs and differentiated neural cells (neurons and astrocytes). We detected all subtypes in neurons but not in astrocytes. In NPCs, CaV3.1 was the dominant subtype, whereas CaV3.2 was weakly expressed, and CaV3.3 was not detected. Next, we determined CaV3.1 expression levels in the cortex during early brain development. Expression levels of CaV3.1 in the embryonic period were transiently decreased during the perinatal period and increased at postnatal day 11. We then pharmacologically blocked T-type calcium channels to determine the effects in neuronal cells. The blockade of T-type calcium channels reduced cell viability, and induced apoptotic cell death in NPCs but not in differentiated astrocytes. Furthermore, blocking T-type calcium channels rapidly reduced AKT-phosphorylation (Ser473) and $GSK3{\beta}$-phosphorylation (Ser9). Our results suggest that T-type calcium channels play essential roles in maintaining NPC viability, and T-type calcium channel blockers are toxic to embryonic neural cells, and may potentially be responsible for neurodevelopmental disorders.

• Food Science of Animal Resources
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• v.21 no.4
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• pp.285-291
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• 2001

The experiment was conducted to study the effect of activated charcoal on the fatty acid composition, meat color and mineral contents of chicken meat with 48 birds of broiler for 6 weeks by adding 0%, 0.6%, 0.9% and 1.2% of activated charcoal to broiler diet. The activated charcoal level of 0.6% and 0.9% increased significantly oleic acid and arachidonic acid contents compared to control(p<0.05). L*, a*, b* values were not influenced by the added level of activated charcoal, however and a* value of different parts of chicken meat was significantly different(p<0.05). The activated charcoal diet increased significantly(p<0.05) the calcium, magnesium and phosphorus content of chicken meat, and tended to increase total mineral contents. In conclusion the activated charcoal supplements to chicken diet increased oleic acid, arachidonic acid the total mineral contents.