• Title/Summary/Keyword: cell surface protein

Search Result 462, Processing Time 0.022 seconds

LAMP-3 (Lysosome-Associated Membrane Protein 3) Promotes the Intracellular Proliferation of Salmonella typhimurium

  • Lee, Eun-Ju;Park, Kwan-Sik;Jeon, In-Sook;Cho, Jae-Woon;Lee, Sang-Jeon;Choy, Hyun E.;Song, Ki-Duk;Lee, Hak-Kyo;Choi, Joong-Kook
    • Molecules and Cells
    • /
    • v.39 no.7
    • /
    • pp.566-572
    • /
    • 2016
  • Lysosomes are cellular organelles containing diverse classes of catabolic enzymes that are implicated in diverse cellular processes including phagocytosis, autophagy, lipid transport, and aging. Lysosome-associated membrane proteins (LAMP-1 and LAMP-2) are major glycoproteins important for maintaining lysosomal integrity, pH, and catabolism. LAMP-1 and LAMP-2 are constitutively expressed in Salmonella-infected cells and are recruited to Salmonella-containing vacuoles (SCVs) as well as Salmonella- induced filaments (Sifs) that promote the survival and proliferation of the Salmonella. LAMP-3, also known as DC-LAMP/CD208, is a member of the LAMP family of proteins, but its role during Salmonella infection remains unclear. DNA microarray analysis identified LAMP-3 as one of the genes responding to LPS stimulation in THP-1 macrophage cells. Subsequent analyses reveal that LPS and Salmonella induced the expression of LAMP-3 at both the transcriptional and translational levels. Confocal Super resolution N-SIM imaging revealed that LAMP-3, like LAMP-2, shifts its localization from the cell surface to alongside Salmonella. Knockdown of LAMP-3 by specific siRNAs decreased the number of Salmonella recovered from the infected cells. Therefore, we conclude that LAMP-3 is induced by Salmonella infection and recruited to the Salmonella pathogen for intracellular proliferation.

Fabrication of PHBV/Keratin Composite Nanofibrous Mats for Biomedical Applications

  • Yuan, Jiang;Xing, Zhi-Cai;Park, Suk-Woo;Geng, Jia;Kang, Inn-Kyu;Yuan, Jiang;Shen, Jian;Meng, Wan;Shim, Kyoung-Jin;Han, In-Suk;Kim, Jung-Chul
    • Macromolecular Research
    • /
    • v.17 no.11
    • /
    • pp.850-855
    • /
    • 2009
  • Keratin is an important protein used in wound healing and tissue recovery. In this study, keratin was modified chemically with iodoacetic acid (IAA) to enhance its solubility in organic solvent. Poly(hydroxybutylate-co-hydroxyvalerate) (PHBV) and modified keratin were dissolved in hexafluoroisopropanol (HFIP) and electrospun to produce nanofibrous mats. The resulting mats were surface-characterized by ATR-FTIR, field-emission scanning electron microscopy (FE-SEM) and electron spectroscopy for chemical analysis (ESCA). The pure m-keratin mat was cross-linked with glutaraldehyde vapor to make it insoluble in water. The biodegradation test in vitro showed that the mats could be biodegraded by PHB depolymerase and trypsin aqueous solution. The results of the cell adhesion experiment showed that the NIH 3T3 cells adhered more to the PHBV/m-keratin nanofibrous mats than the PHBV film. The BrdU assay showed that the keratin and PHBV/m-keratin nanofibrous mats could accelerate the proliferation of fibroblast cells compared to the PHBV nanofibrous mats.

Effects of antibacterial mouth rinses on multiple oral biofilms model (구강세정제가 다중 구강 바이오필름 모델에 미치는 영향)

  • Soo-Kyung Jun;Young-Suk Choi
    • Journal of Korean society of Dental Hygiene
    • /
    • v.23 no.4
    • /
    • pp.209-218
    • /
    • 2023
  • Objectives: To confirm the antibacterial effects of each mouth rinse on multiple oral biofilms in vitro. Methods: The antibacterial effects of different mouth rinses were examined by ATP and counted colony forming units (CFU). Preformed oral biofilms on saliva coated hydroxyapatite (sHA) disks were treated with essential oil and saline; then, the multiple oral biofilms were observed by Scanning electron microscope (SEM). RNA sequencing analysis was performed on total RNA isolated from old biofilms of P. intermedia ATCC 49046. Results: In the CFU measured result compared to controls, preformed multiple oral biofilms were reduced from a low of 39.0% to 95.7% (p<0.05). The size of bacterial cells changed after treatment with the essential oil, and some of the cells ruptured into small pieces of cell debris. Gene expression in P. intermedia ATCC 49046 significantly altered in RNA transcribed and protein translated genes after exposure to essential oil. Conclusions: Mouth rinse solutions with different ingredients had different antibacterial effects and may alter surface structure and gene expression as determined by RNA sequencing.

Parkin Interacts with the PDZ Domain of Multi-PDZ Domain Protein MUPP1 (Parkin과 Multi-PDZ Domain Protein (MUPP1) 단백질 간의 PDZ 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
    • /
    • v.24 no.8
    • /
    • pp.820-826
    • /
    • 2014
  • The localization to specific subcellular sites and the regulation of cell surface receptors and channels are crucial for proper functioning. Postsynaptic density-95/Disks large/Zonula occludens-1 (PDZ)-domain is involved in recognition of and interaction between various proteins, by which the localization and the regulation are mediated. Multi-PDZ domain protein 1 (MUPP1) contains 13 PDZ domains. MUPP1 serves a scaffolding function for structure proteins and signaling proteins, but the mechanism how MUPP1 is stabilized and signalized has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and Parkin. Parkin is an E3 ubiquitin ligase. Loss-of-function mutations of Parkin gene are known to cause an autosomal recessive juvenile parkinsonism. Parkin bound to the $12^{th}$ PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of Parkin has a type II PDZ-association motif, which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, Parkin co-localized with MUPP1. When co-expressed with ubiquitin in HEK-293T cells, MUPP1 has been strongly ubiquitinated by Parkin. These findings collectively suggest that MUPP1 is a novel substrate of Parkin and its function or stability could be modulated by Parkin-mediated ubiquitination.

Immunomodulatory effect of bee pollen extract in macrophage cells (꿀벌 꽃가루 열수 추출물의 큰포식세포 면역활성 효과)

  • Kim, Yi-Eun;Cho, Eun-Ji;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
    • /
    • v.50 no.4
    • /
    • pp.437-443
    • /
    • 2018
  • Activation of macrophages plays an important role in the host-immune system. In this study, we investigated the functional roles and related signaling mechanism of hot-water extracts of bee pollen (BPW) in RAW 264.7 macrophages. Since BPW did not exert cytotoxicity at concentrations ranging from 62.5 to $250{\mu}g/mL$ in macrophage cells, a concentration of $250{\mu}g/mL$ was used as the maximum dose of BPW throughout subsequent experiments. BPW increased inducible nitric oxide synthase-mediated nitric oxide production in a concentration-dependent manner. Additionally, BPW was found to induce macrophage activation by augmenting the expression of cell surface molecules (cluster of differentiation; CD80/86, and major histocompatibility complex; MHC class I/II) and production of pro-inflammatory cytokines (tumor necrosis $factor-{\alpha}$, interleukin-6, and $IL-1{\beta}$) through mitogen-activated protein kinase and nuclear $factor-{\kappa}B$ signaling pathways in RAW 264.7 macrophages. Taken together, our results indicate that BPW could potentially be used as an immunomodulatory agent.

Development of PCR assay for the detection of Brucella spp in bovine semen (종모우 정액중 Brucella균 신속 검출을 위한 PCR기법 개발)

  • Jung, Suck-chan;Jung, Byeong-yeal;Woo, Seong-ryong;Cho, Dong-hee;Kim, Jong-yeom;Kim, Woo-taek;Lee, Jung-mi;Park, Yong-ho;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
    • /
    • v.38 no.2
    • /
    • pp.345-352
    • /
    • 1998
  • The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.

  • PDF

Effects of Red-Koji Fermented Scutellariae Radix Extracts on Lipopolysaccharide-induced Rat Acute Lung Injury (홍국발효 황금이 Lipopolysaccharide 유발 급성 폐손상에 미치는 영향)

  • Kim, Koang Lok;Kwon, Kyoung Man;Yun, Yong Jae;Lee, Young Jun;Park, Dong Il;Kim, Jong Dae;Jung, Tae Young
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.26 no.6
    • /
    • pp.874-885
    • /
    • 2012
  • In the present study, the possibility of whether the pharmacological effects of Scutellariae Radix Aqueous Extracts(SR) were favorably changed by report that lipopolysaccharide(LPS)-induced rat acute lung injury was treated with Red-Koji(Monascus purpureus 12002) fermentation. Three different dosages of Red-Koji fermented SR extract(fSR), 125, 250 and 500 mg/kg were orally administered once a day for 28 days before LPS(Escherichia coli 0111:B4) treatments, and then 5 hours after LPS treatment(500 ${\mu}g$/head, intra trachea instillation), all rats were sacrificed. Changes in the body weights, lung weights, pulmonary transcapillary albumin transit, arterial gas parameters(pH, $PaO_2$ and $PaCO_2$) bronchoalveolar lavage fluid(BALF) protein, lactate dehydrogenase(LDH) and proinflammatory cytokine tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin-$1{\beta}$(IL-$1{\beta}$) contents, total cell numbers, neutrophil and alveolar macrophage ratios, lung malondialdehyde(MDA), myeloperoxidase(MPO), proinflammatory cytokine TNF-${\alpha}$ and IL-$1{\beta}$ contents were observed with histopathology of the lung, changes on luminal surface of alveolus(LSA), thickness of alveolar septum, number of polymorphonuclear neutrophils(PMNs). As results of LPS-injection, dramatical increases in lung weights, pulmonary transcapillary albumin transit increases in $PaCO_2$, decreases in pH of arterial blood and $PaO_2$, increases of BALF protein, LDH, TNF-${\alpha}$ and IL-$1{\beta}$ contents, total cells, neutrophil and alveolar macrophage ratios, lung MDA, MPO, TNF-${\alpha}$ and IL-$1{\beta}$ contents increases were detected with decreases in LSA and increases of alveolar septum and PMNs numbers, respectively as compared with intact control. Especially fSR 125 mg/kg showed quite similar favorable effects on the LPS-induced acute lung injuries as compared with 60 mg/kg of ${\alpha}$-lipoic acid and 250 mg/kg of SR. The results suggest that over 125 mg/kg of fSR extracts showed favorable effects on the LPS-induced acute lung injury mediated by their antioxidant and anti-inflammatory effects. Moreover, increases of the pharmacological effects of SR on LPS-induced acute lung injury were observed by Red-Koji fermentation in this study, at least 2-fold higher.

Engraftment of Intraperitoneally Injected Bone Marrow Cells to Newborn Mice Injected with an Angiogenesis Inhibitor (혈관생성 억제제를 주사한 마우스 모델에서의 골수 세포의 복강 내 주입 후 생착)

  • Cho, Su-Jin;Ju, Sun-Young;Woo, So-Youn;Kang, Hyoung-Jin;Ahn, Hyo-Seop;Ryu, Kyung-Ha;Park, Eun-Ae
    • Neonatal Medicine
    • /
    • v.15 no.1
    • /
    • pp.22-31
    • /
    • 2008
  • Purpose : Bronchopulmonary dysplasia (BPD) is characterized by arrested vascular and alveolar growth in the premature lung. Considering the consequences of arrested lung growth, the idea of administering bone marrow cells to enhance the inborn repair mechanism is promising as this may reduce the morbidity and mortality of BPD. We followed enhanced green fluorescent protein (EGFP)-labeled bone marrow cells (BMC) injected intraperitoneally into non-EGFP mice in order to determine their fate after transplantation. Methods : An angiogenesis inhibitor, SU1498, was injected subcutaneously on day 3 in non-EGFP C57BL/6 newborn mice to create a model of arrested alveolar development. On the following day, $1{\times}10^6$ BMCs isolated from major histocompatibility complex (MHC)- matched syngenic EGFP mice were injected intraperitoneally to non-EGFP BPD mice. Morphometric analysis, immunostaining, and confocal microscopy were performed to determine the fate of EGFP-positive stem cells in the injured lung. Results : SU1498 injection reduced alveolar surface area and mean alveolar volume in newborn mice. BMC injection resulted in recovery of lung structure comparable to controls. EGFP-positive BMCs were identified in the lungs of the recipient mice after intraperitoneal injection. The injected EGFP cells were co-stained with endothelial and epithelial cells of the developing lung as determined by confocal microscopy. Conclusion : Our results illustrated that EGFP-positive BMCs engrafted and trans-differentiated into epithelial and endothelial cells after intraperitoneal injection in a mouse model of arrested alveolar development.

Effects of Lonicerae Flos Extracts on LPS-induced Acute Lung Injury (금은화가 LPS로 유발된 급성 폐 손상에 미치는 영향)

  • Yi, Chang-Geon;Choi, Hae-Yun;Park, Mee-Yeon;Kim, Jong-Dae
    • Journal of Society of Preventive Korean Medicine
    • /
    • v.15 no.1
    • /
    • pp.49-69
    • /
    • 2011
  • Objective : The object of this study was to observe the effects of Lonicerae Flos (LF) aqueous extracts on lipopolysaccharide (LPS)-induced rat acute lung injury. Method : Five different dosages of LF extracts were orally administered once a day for 28 days before LPS treatments, and then all rats were sacrificed after 5 hour-treatment of LPS. Eight groups of 16 rats each were used in the present study. The following parameters caused by LPS treatment were observed ; body weights, lung weights, pulmonary transcapillary albumin transit, arterial gas parameters (pH, $PaO_2$ and $PaCO_2$) bronchoalveolar lavage fluid (BALF) protein lactate dehydrogenase (LDH), and proinflammatory cytokines tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) contents, total cell numbers, neutrophil and alveolar macrophage ratios, lung malondialdehyde (MDA), myeloperoxidase (MPO), proinflammatory cytokines TNF-${\alpha}$ and IL-$1{\beta}$ contents. In addition, the histopathologic changes were observed in the lung in terms of luminal surface of alveolus, thickness of alveolar septum, number of polymorphonuclear neutrophils. Result : As results of LPS-injection, dramatical increases in lung weights, pulmonary transcapillary albumin transit increases, increases in $PaCO_2$, decreases in pH of arterial blood and $PaO_2$, increases of BALF protein, LDH, TNF-${\alpha}$ and IL-$1{\beta}$ contents, total cells, neutrophil and alveolar macrophage ratios, TNF-${\alpha}$ and IL-$1{\beta}$ contents increases were detected with decreases in LSA and increases of alveolar septum and PMNs numbers, respectively as compared with intact control. These are means that acute lung injuries (resembling acute respiratory distress syndrome) are induced by treatment of LPS mediated by inflammatory responses, oxidative stress and related lipid peroxidation in the present study. However, these LPS-induced acute lung injuries were inhibited by 28 days continuous pretreatment of 250 and 500mg/kg of LF extracts. Because of lower three dosages of LF treated groups, 31.25 and 62.5 and 125mg/kg did not showed any favorable effects as compared with LPS control, the effective dosages of LF in LPS-induced acute lung injuries in the present study, is considered as about 125mg/kg. The effects of 250mg/kg of LF extracts showed almost similar effects with ${\alpha}$-lipoic acid 60mg/kg in preventing LPS-induced acute lung injuries. Conclusion : It seems that LF play a role in protecting the acute respiratory distress syndrome caused by LPS.

Effect of Moisture Content on Physical and Chemical Characteristics of Italian Ryegrass Cube (수분 함량이 이탈리안 라이그라스 큐브의 물리적 및 화학적 성상에 미치는 영향)

  • Moon, Byeong Heoun;Park, Hyung Soo;Shin, Jong Seo;Park, Byeong Ki;Kim, Jong Geun
    • Journal of The Korean Society of Grassland and Forage Science
    • /
    • v.36 no.1
    • /
    • pp.34-40
    • /
    • 2016
  • The objective of this study was to determine the effect of moisture content on the physical and chemical characteristics of Italian ryegrass cube. Cube quality according to moisture contents (15, 20, 25, and 30%) was determined. Cubes made with 15 to 20% moisture showed a little cracks. But, the amount of powder generate from these cubes were lower by 10 to 16% compared to other cubes made with 25 to 30% moisture contents. The highest hardness at 159 kg/f was obtained when the cube was made with 15% moisture content and the lowest was 70 kg/f when the cube was made with 30% moisture content. The electrical loading and surface temperature were increased when moisture content was decreased. The chemical compositions of cube were differ from those of raw materials. Crude protein (CP) and ether extract (EE) contents were increased after cubing works. However, crude fiber (CF), acid detergent fiber (ADF), and neutral detergent fiber (NDF) contents were decreased after cubing. The crude ash content was not significantly (p > 0.05) different between raw material and cube. Higher moisture content resulted in higher crude protein content. However, crude fiber and crude ash content were not significantly (p > 0.05) different between each other. The contents of ADF and NDF were the lowest in cubes made with 30% moisture content. Our results suggest that the proper moisture content of Italian ryegrass cubing is recommended to be 15 to 20% and that cubing works should help increase forage quality.