Adhesion events of monocytes represent an important step in inflammatory responses induced by chemokines. The β1-integrin CD29 is a major adhesion molecule regulating leukocyte migration and extravasation. Although several adhesion molecules have been known as regulators of CD29, the molecular interactions between CD29 and its regulatory adhesion molecules (such as CD98 and CD147) have not been fully elucidated. Therefore, in this study, we examined whether these molecules are functionally, biochemically, and cell-biologically associated using monocytic U937 cells treated with aggregation-stimulating and blocking antibodies, as well as enzyme inhibitors. The surface levels of CD29, CD98, and CD147 (but not CD43, CD44, and CD82) were increased. The activation of CD29, CD98, and CD147 by ligation of them with aggregation-activating antibodies triggered the induction of cell-cell adhesion, and sensitivity to various enzyme inhibitors and aggregation-blocking antibodies was similar for CD29-, CD98-, and CD147-induced U937 cell aggregation. Molecular association between these molecules and the actin cytoskeleton was confirmed by confocal microscopy and immunoprecipitation. These results strongly suggest that CD29 might be modulated by its biochemical and cellular regulators, including CD98 and CD147, via the actin cytoskeleton.
Actin Cytoskeleton ; Antibodies ; Antibodies, Blocking ; Chemokines ; Enzyme Inhibitors ; Immunoprecipitation ; Leukocytes ; Ligation ; Microscopy, Confocal ; Monocytes ; U937 Cells*

No abstract available.
Acrocephalosyndactylia*

The actin cytoskeleton plays an important role in macrophage-mediated inflammatory responses by modulating the activation of Src and subsequently inducing nuclear factor (NF)-kappaB translocation. In spite of its critical functions, few papers have examined how the actin cytoskeleton can be regulated by the activation of toll-like receptor (TLR). Therefore, in this study, we further characterized the biological value of the actin cytoskeleton in the functional activation of macrophages using an actin cytoskeleton disruptor, cytochalasin B (Cyto B), and explored the actin cytoskeleton's involvement in morphological changes, cellular attachment, and signaling events. Cyto B strongly suppressed the TLR4-mediated mRNA expression of inflammatory genes such as cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-alpha, and inducible nitric oxide (iNOS), without altering cell viability. This compound also strongly suppressed the morphological changes induced by lipopolysaccharide (LPS), a TLR4 ligand. Cyto B also remarkably suppressed NO production under non-adherent conditions but not in an adherent environment. Cyto B did not block the co-localization between surface glycoprotein myeloid differentiation protein-2 (MD2), a LPS signaling glycoprotein, and the actin cytoskeleton under LPS conditions. Interestingly, Cyto B and PP2, a Src inhibitor, enhanced the phagocytic uptake of fluorescein isothiocyanate (FITC)-dextran. Finally, it was found that Cyto B blocked the phosphorylation of vasodilator-stimulated phosphoprotein (VASP) at 1 min and the phosphorylation of heat shock protein 27 (HSP27) at 5 min. Therefore, our data suggest that the actin cytoskeleton may be one of the key components involved in the control of TLR4-mediated inflammatory responses in macrophages.
Actin Cytoskeleton ; Actins ; Cell Survival ; Cytochalasin B* ; Fluorescein ; Glycoproteins ; HSP27 Heat-Shock Proteins ; Inflammation ; Macrophages ; Membrane Glycoproteins ; Nitric Oxide ; Phosphorylation ; Prostaglandin-Endoperoxide Synthases ; RNA, Messenger ; Toll-Like Receptors ; Tumor Necrosis Factor-alpha

Beauvericin (BEA), a cyclic hexadepsipeptide produced by the fungus Beauveria bassiana, is known to have anti-cancer, anti-inflammatory, and anti-microbial actions. However, how BEA suppresses macrophage-induced inflammatory responses has not been fully elucidated. In this study, we explored the anti-inflammatory properties of BEA and the underlying molecular mechanisms using lipopolysaccharide (LPS)-treated macrophage-like RAW264.7 cells. Levels of nitric oxide (NO), mRNA levels of transcription factors and the inflammatory genes inducible NO synthase (iNOS) and interleukin (IL)-1, and protein levels of activated intracellular signaling molecules were determined by Griess assay, semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter gene assay, and immunoblotting analysis. BEA dose-dependently blocked the production of NO in LPS-treated RAW264.7 cells without inducing cell cytotoxicity. BEA also prevented LPS-triggered morphological changes. This compound significantly inhibited nuclear translocation of the NF-κB subunits p65 and p50. Luciferase reporter gene assays demonstrated that BEA suppresses MyD88-dependent NF-κB activation. By analyzing upstream signaling events for NF-κB activation and overexpressing Src and Syk, these two enzymes were revealed to be targets of BEA. Together, these results suggest that BEA suppresses NF-κB-dependent inflammatory responses by suppressing both Src and Syk.
Beauveria ; Fungi ; Genes, Reporter ; Immunoblotting ; Interleukins ; Luciferases ; Macrophages* ; Nitric Oxide ; Nitric Oxide Synthase ; RNA, Messenger ; Transcription Factors

Flavonoids are plant pigments that have been demonstrated to exert various pharmacological effects including anti-cancer, anti-diabetic, anti-atherosclerotic, anti-bacterial, and anti-inflammatory activities. However, the molecular mechanisms in terms of exact target proteins of flavonoids are not fully elucidated yet. In this study, we aimed to evaluate the anti-inflammatory mechanism of scutellarein (SCT), a flavonoid isolated from Erigeron breviscapus, Clerodendrum phlomidis and Oroxylum indicum Vent that have been traditionally used to treat various inflammatory diseases in China and Brazil. For this purpose, a nitric oxide (NO) assay, polymerase chain reaction (PCR), nuclear fractionation, immunoblot analysis, a kinase assay, and an overexpression strategy were employed. Scutellarein significantly inhibited NO production in a dose-dependent manner and reduced the mRNA expression levels of inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-alpha in lipopolysaccharide (LPS)-activated RAW264.7 cells. In addition, SCT also dampened nuclear factor (NF)-kappaB-driven expression of a luciferase reporter gene upon transfection of a TIR-domain-containing adapter-inducing interferon-beta (TRIF) construct into Human embryonic kidney 293 (HEK 293) cells; similarly, NF-kappa B nuclear translocation was inhibited by SCT. Moreover, the phosphorylation levels of various upstream signaling enzymes involved in NF-kappaB activation were decreased by SCT treatment in LPS-treated RAW264.7 cells. Finally, SCT strongly inhibited Src kinase activity and also inhibited the autophosphorylation of overexpressed Src. Therefore, our data suggest that SCT can block the inflammatory response by directly inhibiting Src kinase activity linked to NF-kappaB activation.
Brazil ; China ; Clerodendrum ; Erigeron ; Flavonoids ; Genes, Reporter ; Humans ; Interferon-beta ; Kidney ; Luciferases ; Macrophages ; NF-kappa B ; Nitric Oxide ; Nitric Oxide Synthase ; Phosphorylation ; Phosphotransferases* ; Plants ; Polymerase Chain Reaction ; RNA, Messenger ; Transfection ; Tumor Necrosis Factor-alpha

Since previous studies have reported that hydroquinone (HQ) exerted immunosuppressive and anti-inflammatory activity, various HQ derivatives have been synthesized and their biological activities investigated. In this study, we explored the anti-inflammatory activity of JS-III-49, a novel HQ derivative, in macrophage-mediated inflammatory responses. JS-III-49 suppressed the production of the inflammatory mediators nitric oxide (NO) and prostaglandin E2 (PGE2) and down-regulated the mRNA expression of the inflammatory enzymes cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) as well as the expression of the pro-inflammatory cytokines interleukin-6 (IL-6) and IL-1b without cytotoxicity in LPS-stimulated RAW264.7 cells. JS-III-49 inhibited nuclear translocation of the NF-kB transcription factors p65 and p50 by directly targeting Akt, an upstream kinase of the NF-kB pathway, in LPS-stimulated RAW264.7 cells. However, JS-III-49 did not directly inhibit the kinase activities of Src and Syk, which are upstream kinases of Akt, in LPS-stimulated RAW264.7 cells. Moreover, JS-III-49 suppressed the nuclear translocation of c-Fos, one of the components of AP-1, by specifically targeting p38, an upstream mitogen-activated protein kinase (MAPK) in the AP-1 pathway in LPS-stimulated RAW264.7 cells. These results suggest that JS-III-49 plays an anti-inflammatory role in LPS-stimulated macrophages by targeting Akt and p38 in the NF-kB and AP-1 pathways, respectively.
Cyclooxygenase 2 ; Cytokines ; Dinoprostone ; Interleukin-6 ; Macrophages ; NF-kappa B ; Nitric Oxide ; Nitric Oxide Synthase Type II ; Phosphotransferases ; Protein Kinases ; RNA, Messenger ; Transcription Factor AP-1 ; Transcription Factors

No abstract available.
Cystectomy* ; Quality of Life* ; Urinary Diversion*

PURPOSE: We analyzed the oncologic outcome of the malignant transformed benign giant cell tumor of bone. MATERIALS AND METHODS: Between January 2000 and February 2012, 5 cases were referred with suspicious malignant transformation of benign giant cell tumor. No patients underwent radiation therapy. RESULTS: After referral, all patients received the wide excision of the tumor and its' pathologic diagnosis were osteosarcoma. As classified by the location of tumor lesion, 3 cases were located in the distal femur, 1 case was in the distal radius and 1 case was in the proximal femur. The average latent period between diagnosis of benign giant cell tumor and diagnosis of secondary malignant giant cell tumor was 49.2 months. (range, 24-126 months) The mean follow-up period was 21.6 months. There were subsequent local recurrence in 2 cases and 3 patients developed distant metastasis. All patients with lung metastasis were dead. CONCLUSION: Malignant transformation of benign giant cell tumor of bone can be occurred within 5 years. Therefore, when benign giant cell tumor suspicious malignant transformation, it is necessary to do more aggressive treatment.
Femur ; Follow-Up Studies ; Giant Cell Tumor of Bone ; Giant Cell Tumors ; Giant Cells ; Humans ; Lung ; Neoplasm Metastasis ; Osteosarcoma ; Radius ; Recurrence ; Referral and Consultation

BACKGROUND: Defects in apoptotic signaling pathways play important role in tumor initiation, progression, metastasis and resistance to treatment. Several proteins which may promote tumorigenesis by inhibiting apoptosis were identified. The survivin protein is the member of inhibitor of apoptosis protein(IAPs) family which inhibits apoptosis. Unlike other IAPs, it is expressed in during the fetal period but not in adult differentiated tissues. Many reports have stated that survivin is selectively expressed in many cancer cell lines and cancer tissues. We performed immunohistochemical analysis for survivin expression in non-small cell lung cancer to get evaluate its clinical implication. METHODS: Twenty nine surgically resected lung cancers were examined. Immunohistochemical staining were performed by immuno-peroxidase technique using avidin-biotinylated horseradish peroxidase complex in the formalin-fixed, paraffin-embedded tissue 4 µm section. Anti-survivin polyclonal antibody was used for primary antibody and anti-p53 monoclonal antibody was also used to analyze the correlation between survivin and p53 expression. The survivin expression scores were determined by as the sum of the stained area and intensity. RESULTS: Immunohistochemical analysis showed cancer specific expression of survivin in 20 of 29 cases (69.0%). Western blot analysis also showed the selective survivin expression in turmor tissue. There was no correlation between survivin expression and clinicopathological parameters and prognosis. We analyzed the correlated between survivin expression and p53 expression, but found none. CONCLUSION: We confirmed the tumor specific expression of survivin in non-small cell lung cancer But this pression was not correlated with clinical parameters as well as histlogy, tumor stage recurrence, and sur rate. Also it ws not statistically correlated with the expression of p53.
Adult ; Apoptosis ; Blotting, Western ; Carcinogenesis ; Carcinoma, Non-Small-Cell Lung* ; Cell Line ; Horseradish Peroxidase ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins* ; Lung Neoplasms ; Neoplasm Metastasis ; Prognosis ; Recurrence

BACKGROUND: Knowledge of size and morphology of the normal trachea is important for airway management and tracheal reconstruction. Conventional radiography is a simple method used to measure the tracheal diameter, but it is not accurate because of the artifacts related to image magnification and overlapping by the shoulder. The purpose of this study was to provide the normal values of the tracheal size and anatomy in Korean adults using Computerized Tomography. MATERIAL AND METHOD: There were 43 men and 34 women included in this study. They were divided into three age groups(group 1, 20-39 years ; group 2, 40-59 yeas ; groups 3,>or=60 years). The anteroposterior and transverse diameters and cross - sectional areas of the trachea were measured at the level of the thoracic inlet(Level 1) and the aortic arch(Level 2). These values obtained at each level were compared between age groups and sexes. RESULT: In 43 men, the anteroposterior / transverse diameters(mean SD in millimeters) of the trachea at levels 1 and 2 were 19.95+/-2.99 / 17.72+/-2.13 and 19.77+/-2.57 / 18.02+/-2.19, respectively. In 34 women, those values at levels 1 and 2 were 15.56+/-2.12 / 14.18+/-2.07 and 15.35+/-1.82 / 15.00+/-1.60, respectively. At both levels, the anteroposterior and transverse diameters were significantly greater in men than in women (p<0.05). The cross-sectional area of the trachea at levels 1 and 2 were 279.14+/-61.37 / 281.93+/-63.97 mm2 in men and 173.29+/-35.81 / 181.88+/-34.74 in women, respectively. They also showed significantly greater values in men than in women(P<0.05). There was no significant difference in diameters and cross-sectional areas of the trachea between age groups. CONCLUSION: There are significant differences in the internal diameter and cross- sectional area of the trachea between men and women in normal Korean adults, while the age difference was insignificant. We believed CT is a relatively accurate and safe way to measure the internal diameter and cross-sectional areas of the trachea.
Adult* ; Airway Management ; Artifacts ; Female ; Humans ; Male ; Radiography ; Reference Values ; Shoulder ; Tomography, X-Ray Computed ; Trachea*