PKC-mediated N-glycosylation of NOX2 regulates its trafficking to the lipid rafts and ROS-dependent degranulation in human mast cells stimulated with protozoan parasite Trichomonas vaginalis-derived secretory products.

Arim Min,Young Hee Nam,Young-Ah Lee,Kyoung-Ju Song,Myeong Heon Shin 한국당과학회 2012 한국당과학회 학술대회 Vol.2012 No.1

Degranulation in human mast cells contributes to provocation of allergic inflammation and innate immunity to parasites. Trichomonas vaginalis is transmitted by sexual intercourse and causes the acute and chronic allergic inflammation. Although a recent report has shown that mast cells are degranulated in response to T. vaginalis-derived secretory products (TvSP), detailed signaling mechanisms of TvSP- induced mast cell degranulation are not fully understood. We report that N-glycosylation of NOX2 play an important role in surface trafficking of NOX2, which can regulate ROS-dependent degranulation in TvSP- stimulated human mast cells. Stimulation with TvSPinduced NOX2 activation such as p47phox phosphorylation and NOX2 glycosylation, production of intracellular ROS, and release of granular proteins such as histamine and -hexosaminidase via BLT1. Inhibition of ROS generation with NOX2 inhibitors also reduced TvSP-triggered degranulation. Moreover, TvSP promoted trafficking of intracellular NOX2 to the cell surface in a BLT1-dependent manner. Such BLT1-mediated surface migration of NOX2 was dependent upon N-glycosyation status of NOX2. Inhibiting N-glycosylation of NOX2 by N-glycosylation inhibitor tunicamycin reduced surface trafficking of NOX2, ROS generation, and degranulation induced by TvSP. PKC inhibitor prevented TvSP-triggered glycosylation of NOX2, its surface trafficking, ROS generation, degranulation. Finally, disruption of lipid rafts with M □CD prevented glycosylation-dependent surface trafficking of NOX2, ROS generation, and degranulation. These results suggest that PKC-mediated N- glycosylation regulates its trafficking of NOX2 to the lipid rafts, which is required for ROS-dependent degranulation in mast cells induced by T. vaginalis-derived secretory products.

Leukotriene B(4) receptors BLT1 and BLT2 are involved in interleukin-8 production in human neutrophils induced by Trichomonas vaginalis-derived secretory products.

Nam, Young Hee,Min, Arim,Kim, Seong Hoon,Lee, Young Ah,Kim, Kyeong Ah,Song, Kyoung-Ju,Shin, Myeong Heon Birkhäuser 2012 Inflammation research Vol.61 No.2

<P>Trichomonas vaginalis is a flagellated protozoan parasite that causes human trichomoniasis. Although T. vaginalis itself can secrete lipid mediator leukotriene (LT) B(4) leading to neutrophil activation, information regarding the signaling mechanism involved in neutrophil activation induced by T. vaginalis-secreted LTB(4) is limited. We investigated whether LTB(4) contained in the T. vaginalis-derived secretory products (TvSP) is closely involved in interleukin (IL)-8 production in human neutrophils via LTB(4) receptors BLT1 or BLT2.</P>

Visfatin Promotes Wound Healing through the Activation of ERK1/2 and JNK1/2 Pathway

Lee, Byung-Cheol,Song, Jisun,Lee, Arim,Cho, Daeho,Kim, Tae Sung MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.11

<P>Visfatin, a member of the adipokine family, plays an important role in many metabolic and stress responses. The mechanisms underlying the direct therapeutic effects of visfatin on wound healing have not been reported yet. In this study, we examined the effects of visfatin on wound healing in vitro and in vivo. Visfatin enhanced the proliferation and migration of human dermal fibroblasts (HDFs) and keratinocytes the expression of wound healing-related vascular endothelial growth factor (VEGF) in vitro and in vivo. Treatment of HDFs with visfatin induced activation of both extracellular signal-regulated kinases 1 and 2 (ERK1/2) and c-Jun N-terminal kinases 1 and 2 (JNK1/2) in a time-dependent manner. Inhibition of ERK1/2 and JNK1/2 led to a significant decrease in visfatin-induced proliferation and migration of HDFs. Importantly, blocking VEGF with its neutralizing antibodies suppressed the visfatin-induced proliferation and migration of HDFs and human keratinocytes, indicating that visfatin induces the proliferation and migration of HDFs and human keratinocytes via increased VEGF expression. Moreover, visfatin effectively improved wound repair in vivo, which was comparable to the wound healing activity of epidermal growth factor (EGF). Taken together, we demonstrate that visfatin promotes the proliferation and migration of HDFs and human keratinocytes by inducing VEGF expression and can be used as a potential novel therapeutic agent for wound healing.</P>

BLT1-mediated O-glycosylation is involved in CREB and NF-, B –dependent IL-8 production in human mast cell induced by protozoan parasite Trichomonas vaginalis-secretory products

Young Hee Nam,Arim Min,Young-Ah Lee,Kyoung-Ju Song,Myeong Heon Shin 한국당과학회 2012 한국당과학회 학술대회 Vol.2012 No.1

Trichomonas vaginalis is a flagellated protozoan parasite causing vaginal trichomoniasis in women, and can secrete chemotactic lipid mediator LTB4 in their secretory products. However, signaling mechanisms of tissue inflammatory responses by infection with T. vaginalis are not fully understood. Recently, we have recently demonstrated that human mast cells and neutrophils are activated to produce chemokine IL-8 via LTB4 receptor BLT1 in response to T. vaginalis-derived secretory products (TvSP). Here, we report that O-glycosylation is important in BLT1-mediated IL-8 secretion in human mast cells (HMC-1 cell line) induced by TvSP. Incubation of HMC-1 cells with TvSP resulted in marked increase of O-GlcNacylated proteins and up-regulated IL-8 protein secretion. TvSP-induced IL-8 production and O-glycosylation in HMC-1 cells was inhibited by pretreatment with OGT inhibitor or OGT siRNA. Pretreatment of HMC-1 cells with BLT1 antagonist or BLT1 siRNA strongly abolished the stimulatory effects of TvSP on O-GlcNacylation and IL-8 production. Moreover, TvSP-induced phosphorylation of transcription factors NF-B and CREB for IL-8 production was reduced by pretreatment of HMC-1 cells with OGT inhibitor or OGT siRNA. These results suggest that BLT1-mediated O-glycosylation may play an important role in activation of transcription factors CREB and NF-B for IL-8 production in mast cells stimulated with TvSP, which can contribute to mast cell-mediated tissue inflammatory responses in T. vaginalis-infected lesion during human trichomoniasis.

Mutations in gyrA and gyrB among Drug Resistant Mycobacterium Tuberculosis in Korea

( Hee Joo Lee ),( Hwi-Jun Kim ),( Ryeun Heo ),( Cheon-Tae Kim ),( Hee-Jin Kim ),( Jeong-hui Gwon ),( Gicheon Bae ),( Sumi Kang ),( Soul-hee Kim ),( Seungmo Kim ),( Eunseon Kim ),( Arim Song ),( Dea-Se 대한결핵 및 호흡기학회 2021 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.129 No.0

Purpose In 2020 KATRD, we analyzed 65 isolates to see how gyrA and gyrB are associated with 7H9 broth microdilution method (BMD) and Lowenstein-Jensen (L-J) drug susceptibility test (DST) because fluoroquinolones (FLQ) have been recognized as important anti-TB agents. In this study, the aim is to evaluate the correlation between gyrA and gyrB mutations and resistance to FLQ with BMD and L-J DST as a follow up of the previous study. Method Since 2020, we have analyzed 304 INH or RIF mono resistant cases by molecular DST using sequencing for gyrA and gyrB genes of FLQ. MICs were measured by BMD for moxifloxacin (MFX, 0.0625~8.0 μg/mL) and levofloxacin (LFX, 0.0625~8.0 μg/ mL). In L-J DST, concentration of MFX was 1.0 μg/mL, 2.0 μg/mL and LFX was 2.0 μg/mL. Results In gyrA and gyrB sequencing, 270 strains (88.81%) were wild type (WT), 34 strains (11.18%) were mutant type (MT). Among 29 gyrA MT strains, 13 isolates (44.82%) had mutation at D94G, 7 isolates (24.14%) at A90V and 5 isolates (17.24%) at D94A. Among 5 gyrB MT strains, 2 isolates (40%) had D500N mutation. In L-J DST, 100% of gyrA MT strains were resistant to MFX. On the other hand gyrB MT strains, 60% to MFX and 40% to LFX were resistant. In BMD, 93.10% of gyrA MT strains ranged 0.5 ~ 4.0 μg/mL and 60% of gyrB MT strains ranged 0.5 ~ 4.0 μg/mL to MFX. Meanwhile 96.55% of gyrA MT strains ranged 1.0 ~ 8.0 μg/mL and 80% of gyrB MT strains ranged 1.0 ~ 8.0 μg/mL to LFX. Conclusions Most of the mutant isolates had mutations in gyrA and most of mutant type (38.23%) was gyrA_D94G (GAC/GGC). In this study we observed gyrA genes were associated with higher MICs based on 7H9 and L-J DST than gyrB genes. # No.2021R1F1A1061358