• Biomolecules & Therapeutics
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• v.27 no.6
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• pp.522-529
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• 2019

M1/M2 polarization of immune cells including microglia has been well characterized. It mediates detrimental or beneficial roles in neuroinflammatory disorders including cerebral ischemia. We have previously found that sphingosine 1-phospate receptor subtype 1 ($S1P_1$) in post-ischemic brain following transient middle cerebral artery occlusion (tMCAO) can trigger microglial activation, leading to brain damage. Although the link between $S1P_1$ and microglial activation as a pathogenesis in cerebral ischemia had been clearly demonstrated, whether the pathogenic role of $S1P_1$ is associated with its regulation of M1/M2 polarization remains unclear. Thus, this study aimed to determine whether $S1P_1$ was associated with regulation of M1/M2 polarization in post-ischemic brain. Suppressing $S1P_1$ activity with its functional antagonist, AUY954 (5 mg/kg, p.o.), attenuated mRNA upregulation of M1 polarization markers in post-ischemic brain at 1 day and 3 days after tMCAO challenge. Similarly, suppressing $S1P_1$ activity with AUY954 administration inhibited M1-polarizatioin-relevant $NF-{\kappa}B$ activation in post-ischemic brain. Particularly, $NF-{\kappa}B$ activation was observed in activated microglia of post-ischemic brain and markedly attenuated by AUY954, indicating that M1 polarization through $S1P_1$ in post-ischemic brain mainly occurred in activated microglia. Suppressing $S1P_1$ activity with AUY954 also increased mRNA expression levels of M2 polarization markers in post-ischemic brain, further indicating that $S1P_1$ could also influence M2 polarization in post-ischemic brain. Finally, suppressing $S1P_1$ activity decreased phosphorylation of M1-relevant ERK1/2, p38, and JNK MAPKs, but increased phosphorylation of M2-relevant Akt, all of which were downstream pathways following $S1P_1$ activation. Overall, these results revealed $S1P_1$-regulated M1/M2 polarization toward brain damage as a pathogenesis of cerebral ischemia.

• Current Optics and Photonics
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• v.5 no.4
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• pp.351-361
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• 2021

Speckle imaging is capable of dynamic data acquisition at high spatiotemporal resolution, and has played a vital role in the functional study of biological specimens. The presence of various optical scatterers within the tissue causes alteration of speckle contrast. Thus structures like blood vessels can be delineated and quantified. Although laser speckle imaging is frequently used, an optimization process to ensure the maximum speckle contrast has not been available. In this respect, we here report an experimental procedure to optimize speckle contrast via applying different combinations of varying polarization of the illuminating laser light and multiple analyzer angles. Specifically, samples were illuminated by the p-polarization, 45°-polarization, and s-polarization of the incident laser, and speckle images were recorded without and with the analyzer rotated from 0° to 180° (Δ = 30°). Following the baseline imaging of a solid diffuser and a fixed brain sample, laser speckle contrast imaging (LSCI) was successfully performed to visualize in vivo mouse-brain blood flow. For oblique laser illumination, the maximum contrast achieved with p-polarized and s-polarized light was perpendicular to the analyzer's axis. This study demonstrates the optimization process for maximizing the speckle contrast, which can improve blood-flow estimation in vivo.

• The Korean Journal of Physiology and Pharmacology
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• v.28 no.3
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• pp.239-252
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• 2024

Dexmedetomidine displays multiple mechanisms of neuroprotection in ameliorating ischemic brain injury. In this study, we explored the beneficial effects of dexmedetomidine on blood-brain barrier (BBB) integrity and neuroinflammation in cerebral ischemia/reperfusion injury. Sprague-Dawley rats were subjected to middle cerebral artery occlusion (MCAO) for 1.5 h and reperfusion for 24 h to establish a rat model of cerebral ischemia/reperfusion injury. Dexmedetomidine (9 ㎍/kg) was administered to rats 30 min after MCAO through intravenous injection, and SB203580 (a p38 MAPK inhibitor, 200 ㎍/kg) was injected intraperitoneally 30 min before MCAO. Brain damages were evaluated by 2,3,5-triphenyltetrazolium chloride staining, hematoxylin-eosin staining, Nissl staining, and brain water content assessment. BBB permeability was examined by Evans blue staining. Expression levels of claudin-5, zonula occludens-1, occludin, and matrix metalloproteinase-9 (MMP-9) as well as M1/M2 phenotypes-associated markers were assessed using immunofluorescence, RT-qPCR, Western blotting, and gelatin zymography. Enzyme-linked immunosorbent assay was used to examine inflammatory cytokine levels. We found that dexmedetomidine or SB203580 attenuated infarct volume, brain edema, BBB permeability, and neuroinflammation, and promoted M2 microglial polarization after cerebral ischemia/reperfusion injury. Increased MMP-9 activity by ischemia/reperfusion injury was inhibited by dexmedetomidine or SB203580. Dexmedetomidine inhibited the activation of the ERK, JNK, and p38 MAPK pathways. Moreover, activation of JNK or p38 MAPK reversed the protective effects of dexmedetomidine against ischemic brain injury. Overall, dexmedetomidine ameliorated brain injury by alleviating BBB permeability and promoting M2 polarization in experimental cerebral ischemia/reperfusion injury model by inhibiting the activation of JNK and p38 MAPK pathways.

• Molecules and Cells
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• v.44 no.5
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• pp.281-291
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• 2021

Tissue-resident macrophages play an important role in maintaining tissue homeostasis and innate immune defense against invading microbial pathogens. Brain-resident macrophages can be classified into microglia in the brain parenchyma and non-parenchymal brain macrophages, also known as central nervous system-associated or border-associated macrophages, in the brain-circulation interface. Microglia and non-parenchymal brain macrophages, including meningeal, perivascular, and choroid plexus macrophages, are mostly produced during embryonic development, and maintained their population by self-renewal. Microglia have gained much attention for their dual roles in the maintenance of brain homeostasis and the induction of neuroinflammation. In particular, diverse phenotypes of microglia have been increasingly identified under pathological conditions. Single-cell phenotypic analysis revealed that microglia are highly heterogenous and plastic, thus it is difficult to define the status of microglia as M1/M2 or resting/activated state due to complex nature of microglia. Meanwhile, physiological function of non-parenchymal brain macrophages remain to be fully demonstrated. In this review, we have summarized the origin and signatures of brain-resident macrophages and discussed the unique features of microglia, particularly, their phenotypic polarization, diversity of subtypes, and inflammasome responses related to neurodegenerative diseases.

• Molecules and Cells
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• v.43 no.6
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• pp.581-589
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• 2020

Neurons have multiple dendrites and single axon. This neuronal polarity is gradually established during early processes of neuronal differentiation: generation of multiple neurites (stages 1-2); differentiation (stage 3) and maturation (stages 4-5) of an axon and dendrites. In this study, we demonstrated that the neuron-specific n-glycosylated protein NELL2 is important for neuronal polarization and axon growth using cultured rat embryonic hippocampal neurons. Endogenous NELL2 expression was gradually increased in parallel with the progression of developmental stages of hippocampal neurons, and overexpression of NELL2 stimulated neuronal polarization and axon growth. In line with these results, knockdown of NELL2 expression resulted in deterioration of neuronal development, including inhibition of neuronal development progression, decreased axon growth and increased axon branching. Inhibitor against extracellular signal-regulated kinase (ERK) dramatically inhibited NELL2-induced progression of neuronal development and axon growth. These results suggest that NELL2 is an important regulator for the morphological development for neuronal polarization and axon growth.

• Journal of Surface Science and Engineering
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• v.41 no.5
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• pp.181-188
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• 2008

A.C. impedance properties of HA/Ti compound layer coated Ti-30Ta-($3{\sim}15$)Nb alloys have been studied by electrochemical method. Ti-30Ta binary alloys contained 3, 7, 10 and 15 wt% Nb were manufactured by the vacuum furnace system. And then specimen was homogenized at $1000^{\circ}C$ for 24 hrs. The sample was cut and polished for corrosion test and coating. It was coated with HA/Ti compound layer by magnetron sputter. The non-coated and coated morphology of Ti alloy were analyzed by X-ray diffractometer (XRD), energy X-ray dispersive spectroscopy (EDX) and filed emission scanning electron microscope (FE-SEM). The corrosion behaviors were investigated using A.C. impedance test (PARSTAT 2273, USA) in 0.9% NaCl solution at $36.5{\pm}1^{\circ}C$. Ti-30Ta-($3{\sim}15\;wt%$)Nb alloys showed the ${\alpha}+{\beta}$ phase, and $\beta$ phase peak was predominantly appeared in the case of increasingly Nb contents. The microstructures of Ti alloy were transformed from needle-like structure to equiaxed structure as Nb content increased. From the analysis of coating surface, HA/Ti composite surface uniformed coating layer with 750 nm thickness. The growth directions of film were (211), (112), (300) and (202) for HA/Ti composite coating on the surface after heat treatment at $550^{\circ}C$, whereas, the growth direction of film was (110) for Ti coating. The polarization resistance ($R_p$) of HA/Ti composite coated Ti-alloys were higher than those of the Ti and HA coated samples in 0.9% NaCl solution at $36.5{\pm}1^{\circ}C$. Especially, corrosion resistance of Ti-Ta-Nb system increased as Nb content increased.

• Journal of Surface Science and Engineering
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• v.41 no.2
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• pp.69-75
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• 2008

Electrochemical behaviors of surface modified and MC3T3-E1 cell cultured Ti-30Ta alloys have been investigated using various electrochemical methods. The Ti alloys containing Ta were melted by using a vacuum furnace and then homogenized for 6 hrs at $1000^{\circ}C$. MC3T3-E1 cell culture was performed with MC3T3-E1 mouse osteoblasts for 2 days. The microstructures and corrosion resistance were measured using FE-SEM, XRD, EIS and potentiodynamic test in artificial saliva solution at $36.5{\pm}1^{\circ}C$. Ti-Ta alloy showed the martensite structure of ${\alpha}+{\beta}$ phase and micro-structure was changed from lamellar structure to needle-like structure as Ta content increased. Corrosion resistance increased as Ta content increased. Corrosion resistance of cell cultured Ti-Ta alloy increased predominantly in compared with non cell cultured Ti- Ta alloy due to inhibition of the dissolution of metal ion by covered cell. $R_p$ value of MC3T3-E1 cell cultured Ti-40 Ta alloy showed $1.60{\times}10^6{\Omega}cm^2$ which was higher than those of other Ti alloy. Polarization resistance of cell-cultured Ti-Ta alloy increased in compared with non-cell cultured Ti alloy.

• BMB Reports
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• v.52 no.10
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• pp.613-618
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• 2019

Microglial cells are known as the main immune cells in the central nervous system, both regulating its immune response and maintaining its homeostasis. Furthermore, the antioxidant ${\alpha}-lipoic$ acid (LA) is a recognized therapeutic drug for diabetes because it can easily invade the blood-brain barrier. This study investigated the effect of ${\alpha}-LA$ on the inflammatory response in lipopolysaccharide (LPS)-treated BV-2 microglial cells. Our results revealed that ${\alpha}-LA$ significantly attenuated several inflammatory responses in BV-2 microglial cells, including pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ and interleukin (IL)-6, and other cytotoxic molecules, such as nitric oxide and reactive oxygen species. In addition, ${\alpha}-LA$ inhibited the LPS-induced phosphorylation of ERK and p38 and its pharmacological properties were facilitated via the inhibition of the nuclear factor kappa B signaling pathway. Moreover, ${\alpha}-LA$ suppressed the activation of NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasomes, multiprotein complexes consisting of NLRP3 and caspase-1, which are involved in the innate immune response. Finally, ${\alpha}-LA$ decreased the genes accountable for the M1 phenotype, $IL-1{\beta}$ and ICAM1, whereas it increased the genes responsible for the M2 phenotype, MRC1 and ARG1. These findings suggest that ${\alpha}-LA$ alleviates the neuroinflammatory response by regulating microglial polarization.

• Korean Journal of Metals and Materials
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• v.47 no.2
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• pp.129-137
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• 2009

Corrosion behaviors of TiN coated dental casting alloys have been researched by using various electrochemical methods. Three casting alloys (Alloy 1: 63Co-27Cr-5.5Mo, Alloy 2: 63Ni-16Cr-5Mo, Alloy 3: 63Co-30Cr-5Mo) were prepared for fabricating partial denture frameworks with various casting methods; centrifugal casting(CF), high frequency induction casting(HFI) and vacuum pressure casting(VP). The specimens were coated with TiN film by RF-magnetron sputtering method. The corrosion behaviors were investigated using potentiostat (EG&G Co, 263A. USA) in 0.9% NaCl solution at $36.5{\pm}1^{\circ}C$. The corrosion morphologies were analyzed using FE-SEM and EDX. Alloy 1 and Alloy 2 showed the ${\alpha}-Co$ and ${\varepsilon}-Co$ phase on the matrix, and it was disappeared in case of TiN coated Alloy 1 and 2. In the Alloy 3, $Ni_2Cr$ second phases were appeared at matrix. Corrosion potentials of TiN coated alloy were higher than that of non-coated alloy, but current density at passive region of TiN coated alloy was lower than that of non-coated alloy. Pitting corrosion resistances were increased in the order of centrifugal casting, high frequency induction casting and vacuum pressure casting method from cyclic potentiodynamic polarization test.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.5
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• pp.409-415
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• 2000

Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to evaluate the effects of dopamine HCl on the range of the rotatioanl mobility of bulk bilayer structure of the synaptosomal plasma membrane vesicles (SPMV) isolated from whole bovine brain. In a dose-dependent manner, dopamine decreased the anisotropy $({\gamma}),$ limiting anisotropy $({\gamma}{infty})$ and order parameter (S) of DPH in the membranes. These indicate that dopamine increased the rotational mobility of the probe in the neuronal membranes. Cationic 1-[4-(trimethylammonio)-phenyl]-6-phenylhexa-1,3,5-hexatriene (TMA-DPH) and anionic 3-[p-(6-phenyl)-1,3,5-hexatrienyl]-phenylpropionic acid (PRO-DPH) were utilized to examine the range of transbilayer asymmetric rotational mobility of the neuronal membranes. Dopamine had a greater increasing effect on the mobility of the inner monolayer as compared to the outer monolayer of the neuronal membranes. It has been proven that dopamine exhibits a selective rather than nonselective fluidizing effect within the transbilayer domains of the SPMV.