Jae-Dong Lee

Curcumin Suppresses the Induction of Indoleamine 2,3-Dioxygenase by Blocking the Janus-activated Kinase-Protein Kinase Cδ-STAT1 Signaling Pathway in Interferon-γ-stimulated Murine Dendritic Cells

Indoleamine 2,3-dioxygenase (IDO) catalyzes the initial and rate-limiting step in the degradation of tryptophan and is strongly induced in interferon-γ (IFNγ)-stimulated dendritic cells (DCs). IDO has recently been established as a key enzyme in T-cell suppression-mediated immune tolerance to tumors. STAT1 phosphorylation appears to play an important role in the control of IDO expression by IFNγ, but the precise regulatory mechanism remains obscure. Here we present a novel mechanism of IFNγ-induced IDO expression in bone marrow-derived dendritic cells. In addition, we demonstrate that curcumin, an active component of turmeric, significantly inhibited the induction of IDO expression and activity by IFNγ. We found that curcumin suppressed STAT1 activation by directly inhibiting Janus-activated kinase 1/2 and protein kinase Cδ phosphorylation in bone marrow-derived DCs, suppressing the subsequent translocation and binding of STAT1 to the GAS element of the IRF-1 promoter. Coincident with these inhibitory effects on IFNγ-induced IDO expression, curcumin reversed IDO-mediated suppression of T-cell responses. Our results, thus, suggest that down-regulation of IDO in DCs is an important immunomodulatory property of curcumin that may be exploited therapeutically in the control of cancers.

IL-17 increased the production of vascular endothelial growth factor in rheumatoid arthritis synoviocytes

Interleukin-17 (IL-17) is a proinflammatory cytokine that is expressed in the synovium T cells of rheumatoid arthritis (RA). This cytokine is implicated in the inflammation and destruction of the joint. However, the role of IL-17 on the production of vascular endothelial factor (VEGF) important to synovial proliferation has still not been identified. In this study, we investigated the effect on cultured rheumatoid fibroblast-like synoviocytes (FLS) of the IL-17 on the production and expression of VEGF, which play an important role in angiogenesis in rheumatoid synovium. IL-17 increased the production of VEGF dose dependently and the mRNA expression of VEGF. These results suggest that IL-17 might influence angiogenesis in RA by up-regulating the expression of VEGF in rheumatoid FLS.

The novel role of platelet-activating factor in protecting mice against lipopolysaccharide-induced endotoxic shock.

Background Platelet-activating factor (PAF) has been long believed to be associated with many pathophysiological processes during septic shock. Here we present novel activities for PAF in protecting mice against LPS-mediated endotoxic shock. Principal Findings In vivo PAF treatment immediately after LPS challenge markedly improved the survival rate against mortality from endotoxic shock. Administration of PAF prominently attenuated LPS-induced organ injury, including profound hypotension, excessive polymorphonuclear neutrophil infiltration, and severe multiple organ failure. In addition, PAF treatment protects against LPS-induced lymphocytes apoptosis. These protective effects of PAF was correlated with significantly decreases in the production of the inflammatory mediators such as TNF-α, IL-1β, IL-12, and IFN-γ, while increasing production of the anti-inflammatory cytokine IL-10 in vivo and in vitro. Conclusions Taken together, these results suggest that PAF may protect mice against endotoxic shock via a complex mechanism involving modulation of inflammatory and anti-inflammatory mediators.

JNK inhibitor SP600125 promotes the formation of polymerized tubulin, leading to G2/M phase arrest, endoreduplication, and delayed apoptosis.

The JNK inhibitor SP600125 strongly inhibits cell proliferation in many human cancer cells by blocking cell-cycle progression and inducing apoptosis. Despite extensive study, the mechanism by which SP600125 inhibits mitosis-related effects in human leukemia cells remains unclear. We investigated the effects of SP600125 on the inhibition of cell proliferation and the cell cycle, and on microtubule dynamics in vivo and in vitro. Treatment of synchronized leukemia cells with varying concentrations of SP600125 results in significant G2/M cell cycle arrest with elevated p21 levels, phosphorylation of histone H3 within 24 h, and endoreduplication with elevated Cdk2 protein levels after 48 h. SP600125 also induces significant abnormal microtubule dynamics in vivo. High concentrations of SP600125 (200 µM) were required to disorganize microtubule polymerization in vitro. Additionally, SP600125-induced delayed apoptosis and cell death was accompanied by significant poly ADP-ribose polymerase (PARP) cleavage and caspase-3 activity in the late phase (at 72 h). Endoreduplication showed a greater increase in ectopic Bcl-2-expressing U937 cells at 72 h than in wild-type U937 cells without delayed apoptosis. These results indicate that Bcl-2 suppresses apoptosis and SP600125-induced G2/M arrest and endoreduplication. Therefore, we suggest that SP600125 induces mitotic arrest by inducing abnormal spindle microtubule dynamics.

β-Lapachone (LAPA) decreases cell viability and telomerase activity in leukemia cells: suppression of telomerase activity by LAPA.

Up-regulation of telomerase activity is associated with immortalization and unlimited cell division in most cancer cells. Therefore, telomerase represents a particularly attractive target for anticancer therapy. Recent reports have suggested that beta-lapachone (LAPA), the product of the South American Tabebuia avellanedae tree, inhibits growth of tumor cells. However, the underlying relationship between telomerase activity and apoptosis in response to LAPA exposure in leukemia cells remains poorly understood. In this study, we confirmed that LAPA treatment induces direct cytotoxicity in human leukemia cells (U937, K562, HL60, and THP-1) through activation of caspase-3 and subsequent cleavage of poly(ADP-ribose) polymerase. The observed induction of cell death was associated with decreased telomerase activity, which was ascribed to down-regulation of telomerase reverse transcriptase. Additionally, overexpression of anti-apoptotic Bcl-2 could not overcome the induction of apoptosis or the decreased telomerase activity in response to treatment of U937 cells with LAPA. We conclude that LAPA has a direct cytotoxic effect and the loss of telomerase activity in leukemia cells.

Taxonomical Classification and Species-specific Detection of Genus Some Phellinus using Phylotype

This study was carried out to identify the phylogenetic relationship of Phellinus species and to know its distribution by comparing the DNA sequences of internal transcribed spacer regions(ITS1 and IST2) and 5.8S ribosomal DNA (rDNA) repeat unit. The Phellinus species had their specific sequences in IST1 and 2 regions depending on suedes. The comparison of the ITS sequences of standard strains indicated that the sequences of ITS1 were more variable than those of ITS2. Nine strains of the commercial products of Phellinus species used in this study were identified as P. lintues, P. baumii, P. igniarius, and P. pini. Most of commercial species were P. pini and P. baumii, and P. gilvus was not found. Also, P. linteus was only found in form of mycelial culture rather than fruiting body. Moreover, the species-specific primers were designed based on ITS sequence data. Each species-specific primers were bound in P. lintues(ITSF-PL2R), P. baumii(PB1F-ITS4R), P. igniarius(IF1-IR3), P. pini(PF1-PR3), and P. gilvus(GF2-GR4), respectively. These primer sets would be useful fer the detection of specific-species among unidentified Phellinus species rapidly.

RAPD Analysis and Cordycepin Concentration of Hybrided Cordyceps militaris Strains by Mating

The C. militaris belongs to entomopathogenic fungi, which have their specific sequences in internal transcribed spacer regions (ITS1 and ITS2) depending on species. In this study, to identify the phylogenetic relationship of the strain hybrided by mating of C. militaris, we compared DNA sequences of ITS regions and 5.8S ribosomal DNA (rDNA) repeat unit of hybrid strain and its parental strains. The result revealed that hybrid strains are C. militaris species. In addition, cordycepins produced by hybrid strains and other strains of C. militaris were analyzed by HPLC with 20mM of mobile phase and C-18 columns. The result indicated that the strain hybrided by mating produce higher concentration of phytochemical cordycepin than other C. militaris strains.

Application of Species-specific DNA Probe to Field Samples of Alexandrium tamarense (Lebour) Balech

Fluorescent species-specific DNA probe (AT1) of toxic dinoflagellate Arexandrium tamarense was tested on several other species, on comparison of binding activity at different preservatives for fixation of the cells, at different culture age and estimation of cell density by light microscope or epifluorescent microscope using whole cell hybridization. Th AT1 probe specifically bound to Alexandrium tamarense, whereas it did not bind to other phytoplankton, in particular Alexandrium catenella, morphologically similar to Alexandrium tamarense, could not react to AT1 probe. When cells were fixed with all three preservatives, labeling cells of Alexandrium tamarense emitted strong fluorescent signal intensity. In addition, regardless culture days, binding activity with AT1 probe was strong. The tell densities estimated by epifluorescent microscope were than those estimated by light microscope. The enumeration and identifying of Arexandriurn tamarense using DNA probe method will be contributed to a new biotoxin monitoring and prediction system in field.

Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to he grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

Phylogenetic Relationships between the Genus Inonotus and its Related Genera Based on the Nucleotide Sequences of Internal Transcribed Spacers

In this study the ITS1, ITS2 and 5.8S ribosomal DNA sequences from 29 strains of the Genus Inonotus and its related genera were compared with 31 strains obtained from GenBank database. Using the neighbor-joining (NJ) method and most parsimonious analysis the phylogenetic tree was constructed. The hymenochaetales formed no monophyletic group and several non-hymenochaetales appeared as intermingled with the Hymenochaetales. Strains 6, 46, 49, 50, 53, 55 showed no certain affinities within the Hymenochaetales, whereas Inonotus sp. (51) was closely related to Phellinus baumii, and Inonotus sp. (52), and Inonotus glomeratus (10) was related to Phellinus linteus, and Fomes fomentarius (30) was related to Ganoderma lucidum. Inonotus sp. and Phellinus sp. formed no monophyletic groups and a subdivision in the following genera is accepted: Inonotus sp. Phellinus baumii, Phellinus linteus, Phellinus igniarius, Phellinus pini, Hericium erinaceum, Ganoderma lucidum and Sparassis sp. were confirmed and separated genera. The taxonomic status of Inonotus remained uncertain. Eight new combinations are proposed.