We have analyzed the transcriptome of Scolopendra subspinipes mutilans using RNA sequencing and identified severalantimicrobial peptide candidates. Among the peptides, named scolopendrasins, were selected based on the physicochemicalproperties of antimicrobial peptides via an in silico analysis. As a result, we evaluated the antimicrobial activities ofscolopendrasins against Gram positive and negative bacteria including Candida albicans by radial diffusion assay and colonycount assay. We also investigated the cytotoxicity of scolopendrasins through hemolysis assay. We found that the actionof scolopendrasins involved binding to the surface of microorganisms via a specific interaction with lipopolysaccharides,lipoteichoic acid, and peptidoglycans, which are components of the bacterial membrane. These results will provide a basisfor developing therapeutic agents such as peptide antibiotics.

The dynastid beetle Allomyrina dichotoma has been used as a herbal medicine. Recently, we performed de novo RNAsequencing of Allomyrina dichotoma and identified several antimicrobial peptide candidates based on in silico analysis.Among them, cationic antimicrobial peptide, Allomyrinasin, was selected and we assessed the anti-inflammatory activitiesof Allomyrinasin against mouse macrophage Raw264.7 cells. The results showed that Allomyrinasin decreased the nitricoxide production of the lipopolysaccharide-induced Raw264.7 cells. In addition, quantitative RT-PCR, ELISA and Westernblot analysis revealed that Allomyrinasin reduced cytokine expression levels in the Raw264.7 cells. Taken together, thesedata indicated that Allomyrinasin had anti-inflammatory activity in the lipopolysaccharide-induced Raw264.7 cells.

Although the grasshopper Oxya chinensis sinuosa has long been used as a food in Korea, there is little data on itsfunctional effects. Thus we prepared and analyzed total RNA from the whole body of adult Escherichia coli non-immunizedand immunized Oxya chinensis sinuosa strains. Using an Illumina Hiseq sequencer, we generated a total of 66,555 pooledtranscriptome and singletons with and without Escherichia coli immunization, respectively. Then, we performed in silicoanalysis of the Oxya chinensis sinuosa transcriptome, using bioinformatics tools for screening putative antimicrobial peptides(AMPs) and 38 AMPs were finally selected and tested their antimicrobial activity of Gram positive, Gram negative bacteriaand antifungal activity with radial diffusion assay. As a result, 5 out of 38 AMPs showed the highest antimicrobial activityand antifungal activity against microbes and it also evidenced with no hemolytic activity.

Aphis gossypii is considered as one of the most polyphagous aphid species and found throughout the world. This pestinduce plant deformation and transmit viruses by feeding the host plant. In Korea, A. gossypii has been occurred in manycrops, however, the population genetic structure of this pest has not been revealed. In this study, we used eight microsatellitemarkers to analysis the population genetic structure of A. gossypii in Korea. We collected A. gossypii specimens frompepper greenhouse in 2016 (18 sites) and 2017(15 sites). As a result, two genetic cluster (△K=2) showed in structureanalysis. As we compared the cluster result between 2016 and 2017, several populations appeared opposite clustering groupeven the samples were collected in same site and crop. Therefore, more samples are required from different greenhouse(same host) to identify the accurate genetic cluster of province in Korea. From this study, the genetic variation revealedfrom A. gossypii and this information may develop sustainable management of A. gossypii.

The objective of this study was to evaluate the effect of the Allomyrina dichotoma larva (AD) on allergy and inflammation.We examined inhibitory effect of AD on allergic reactions in mast cells (RBL-2H3) activated by Compound 48 / 80and inflammatory response in macrophages (Raw 264.7) activated by LPS. Anti-allergy and anti-inflammatory actions ofAD water extract had no cytotoxicity. At these concentrations AD inhibited ẞ-hexosaminidase, tumor necrosis factor- α(TNF- α), interleukin-4 (IL-4), interleukin-6(IL-6) and cyclooxygenase-2(COX-2). AD also inhibited the expression of inducibleNO synthase (iNOS). AD reduced the release of inflammatory cytokines including IL-4, IL-6, TNF-α, and ẞ-hexosaminidase.These results suggest that AD may be potential anti-allergy and anti-inflammatory therapeutic agent.

We recently reported the in vitro and in vivo antiobesity effects of Tenebrio molitor larvae, a traditional food in manycountries, but it remains unknown how the larvae affect appetite regulation in mice with diet-induced obesity. We hypothesizedthat the extract of T. molitor larvae mediates appetite by regulating neuropeptide expression. We investigated T. molitorlarvae extract's (TME's) effects on anorexigenesis and endoplasmic reticulum (ER) stress–induced orexigenic neuropeptideexpression in the hypothalami of obese mice. Central administration of TME suppressed feeding by down-regulating theexpression of the orexigenic neuropeptides neuropeptide Y and agouti-related protein. T. molitor larvae extract significantlyreduced the expression of ER stress response genes. These results suggest that TME and its bioactive components arepotential therapeutics for obesity and ER stress–driven disease states.

Entomopathogenic fungus is a useful control agent to sucking type insect such as whitefly and aphid. The fungi are influenced by some environmental factors such as relative humidity, temperature and UV and cause slow and fluctuation in pest control efficacy. Especially, UV kills conidia or spores of entomopathogenic fungi and a mycopesticide using fungi has short control period in field. UV intensity changes from season to season. Survival rate of entomopathognic fungi treated may differ from seasons and will show different control efficacy. Therefore, we conducted a study to estimate the persistence of an Isaria javanica isolate, which was already reported as sweet potato whitefly control agent, in potted greenhouse soil planted different crops. The number of survival spore decreased gradually and differ from seasons.

Cariogenic Streptococcus mutans encounters a variety of host defense factors produced in oral cavity. Nitric oxide (NO) and NO-mediated reactive nitrogen species are potential antimicrobials of innate immunity that can threaten the fitness of S. mutans in their ecological niches. Streptococcal strategies to detoxify cytotoxic NO, which allow S. mutans to persist in caries or other environments of the oral cavity, remain unknown. In this study, we directly measured NO consumption rates of S. mutans isolated in Korea. Surprisingly, all S. mutans strains were unable to consume exogenous NO efficiently, while an intracellular parasite Salmonella enterica serovar Typhimurium expressing the NO-metabolizing enzyme flavohemoglobin consumed most of the NO. This result suggested that S. mutans has alternative detoxification systems for tolerating NO-induced nitrosative stresses.

Human β- defensin-2 (hBD-2) is an antimicrobial peptide which is produced by epithelial cells after stimulation with microorganisms or inflammatory mediators. However, little is known as to whether the LPS and nicotine induces the expression of hBD-2 in periodontal l igament (PDL) cells. T he p urpose o f this s tudy was t o investigate t he r oles o f MAPKs pathway o f nicotine a nd L PS-induced hBD-2 expression in PDL cells. The maximal expression of hBD-2 was observed in nicotine 5 mM and LPS 1 μg/ml treated PDL cells. Nicotine and LPS induced the phosphorylation of p38, c-Jun N-terminal kinase 1 and 2 (JNK1/2), and extracellular signal-regulated kinases 1 and 2 (ERK1/2) MAPK. ERK1/2 inhibitor SB203580, p38 inhibitor PD98059, and JNK inhibitor SP600125, blocked the effects of nicotine and LPS on hBD-2 upregulation in PDL cells. These results collectively suggest that hBD-2 is up-regulated in nicotine and LPS-treated PDL cells through activation of p38, ERK and JNK MAPKs pathway. Our data regarding the up-regulation of hBD-2 may help us to understand the antimicrobial mechanism in periodontal disease

Pleurotus citrinopileatus was successfully cultivated and commercially available in Korea. The antioxidant, xanthine oxidase, tyrosinase inhibitory activities and polyphenol contents of fruiting bodies of Pleurotus citrinopileatus extracted with acetone, hot water and methanol (hereinafter referred to Fr. Ace, Fr. HW and Fr. MeOH). The antioxidant activities on β-carotene-linoleic acid in the Fr. Ace, Fr. HW and Fr. MeOH were 96.12%, 94.21% and 96.52%, respectively at the concentration of 20 mg/ml. Xanthine oxidase inhibition activity in the Fr. Ace, Fr. HW and Fr. MeOH were 30.12%, 35.42% and 29.02%, respectively at the concentration of 5 mg/ml. Tyrosinase inhibition activity in the Fr. Ace, Fr. HW and Fr. MeOH were 58.78%, 49.25% and 63.29%, respectively at the concentration of 1.0 mg/ml. Total polyphenol contents in the Fr. Ace, Fr. HW and Fr. MeOH were 18.99 mgGAEs/g, 16.73 mgGAEs/g and 18.66 mgGAEs/g. These experimental results suggested that fruiting bodies of P. citrinopileatus contained good physio-chemical substances for promoting human health.

The antioxidant, xanthine oxidase, tyrosinase inhibitory activities and polyphenol contents of the fruiting bodies of Pleurotus cornucopae extracted with acetone, hot water and methanol (hereinafter referred to Fr. Ace, Fr. HW and Fr. MeOH). The antioxidant activities in the Fr. Ace, Fr. HW and Fr. MeOH were 93.23%, 89.55% and 92.58%, respectively at the concentration of 2.0 mg/ml. Xanthine oxidase inhibition activity in the Fr. Ace, Fr. HW and Fr. MeOH were 45.84%, 46.50% and 45.60%, respectively at the concentration of 5 mg/ml. Tyrosinase inhibition activity in the Fr. Ace, Fr. HW and Fr. MeOH were 52.11%, 50.12% and 55.81%, respectively at the concentration of 1.0 mg/ml. Total polyphenol contents in the Fr. Ace, Fr. HW and Fr. MeOH were 18.99 mgGAEs/ g, 16.73 mgGAEs/g and 18.66 mgGAEs/g. These experimental results suggested that fruiting bodies of P. cornucopae contained good physio-chemical substances for promoting human health.

Al though the changes in tooth morphology and hardness by hydrogen peroxide(H20 z) have been r‘epor‘.ted .‘ the pαr。o야t뻐ec야tive role of heme oxygenase-l(HO-l) against the cytotoxic effects of H202 has not been clarifïed i n human pulp cells ln this st udy. we investigated whether HO-l is involved in Hz0 2-induced cytoLox icity a nd examined the production 0 1' dentin sia lophosphoprotein(DSPP) and other mineralization markers‘ in hllman pu lp cells H202 decreased cell viabi lity, but increased HO-l and DSPP expression in a concentra tion and time dependent manner . Inhibitors of gllanylate cyclase. PI3K, ERK. and p38 MAP kinase blocked H202-indllced cytotoxicity and the expression of HO-l and DSPP mRNAs in pulp cells. These data suggest that the induction of HO-l by H202 in plllp cells plays a protective role against the cytotoxic effects 0 1' HzOz and stimulates DSPP expression‘ reslllting in prematllre odontoblast dilTerentiation throllgh pathways that involve cGMP‘ p38. and ERK.

Al t hough substance P(SP) , a potent pro- inflammatory peptide, is involved in inflammation and immune responses‘ t he eff'ect of SP on t he expression of macrophage inflammatory protein 3a (MIP- 3α CCL20) in periodontal liga ment(PDL) cell s a re unknown, Equally as enigmatic is the link between SP, t he stress protein heme oxygenase- l(HO-l) ‘ and CCL20 procluction, We investigated whether SP induces the release of chemokine CCL20 from immortal ized PDL(IPDL) ceJJ s‘ and fur ther c l a꺼 SP mediated pathways, We also examined the relationship between HO-l a ncl CCL20 by t reating PDL cells with SP, Incubating IPDL cells with SP increased expression of CCL20 mRNA a nd CCL20 protein in a dose-time dependent manner Highly selective p38 and ERKl/2 inhibitors abrogated SP-induced expression of CCL20 in IPDL cell s, SP is a lso responsible for ini t iating phosphorylation of I/C B, degradation of Iκ B‘ ancl activat ion of NF'-/C B, SP induced expression of HO-l in both a concentration- and time-dependent man nel ‘ and CCL20 refl ected s imilar patterns, The inductive effects o[ SP on HO- l and CCL20 wer e enhanced by HO- j inducer hemin and the membrane-permeable cGMP analog 8-bromo-cGMP, Conversely, this pathway was inJübited by t he 1-10난 inhi bitor zinc protoporphyrin IX(ZnPP IX) and the selective inl뼈itor of guanylate cyc1ase‘ lH-[l , 2, 4Joxad iazole[4‘ 3-aJquinoxal in-l-one (ODQ) , We report herein the pathway that connects SP along with other modulators 。f neuroimmunoregulationto the induction of HO-l and t he inflammatory mediator MIP-3a /CCL20 in IPDL cell s‘ which play an important role in the development 01' periodontitis or inflamrnation during orthodontic tooth movem

1'0 determine Lhe ll1echanism of cell c1eath incluced by iron chelators. we explored the pathways of the three structurally relatecl ll1 itogen-activatecl protein(MAP) kinase subfami li esduring iron cbelator- inclucecl apoptosis ancl differentiation of oral precancerous ancl cancel‘ cells. The iron chelator c1 eferoxamine(DFO) exertecl potent timeancl c1ose-c1epenclent inhi bitory effects on the growth of IHOK and HN4 cells The major mechanism of growth inhibition following DFO treatment was fOllncl to be apoptosis incluction. as assessecl by annexin V-FITC staining. cell cycle analysis‘ DNA lacldering, a ncl Hoechst staining. We report that DF'O s trongly activates the p38 MAP kinase and extracell ular signal- regu lated kinase(ERK). but c10es not activate the c-Jun N-terminal kin ase/ stl않s-activaLecl protein kinase(JNK/8APK) . Of the three MAP kinase blockers usecl‘ the selective p38 MAP kinase inhibitor 8ß203580 ancl ERK inhibitor PD98059 protected oral premaIignant ancl malignant cells againsL iron chelator- nclllced cell death. which incl icates that the p38 MAP kinase serves as a major mecliator 01' apoptos is induced by this iron chelator DFO also evoked the release of cytochrome c from mitochondria, and incluced the activation of caspase-3 ancl caspase-8 in oral cancer cells, which suggests that apoptosis occurs via the mi tochoncl ri on - mecl iaLed pathway. DFO enhanced the expression of Bax in IHOK ancl HN4 cells. consistent witll thei r p53 status Moreover. DFO downregulatecl the expression 01' Bcl-2 in oral cancer cells. which suggests that DFO- incluced apoptos is 01' oraJ cancer and precancerous cells may be mediatecl by an increase in the ratio of pro-apoptotic to anti-apoptotic proteins. ln terestingJy, trcatmcnt 01' IHOK ancl HN4 cel ls with 8B203580 abolishecl cytochrome c release‘ as wel l as the activation of caspase-3 and caspase-8. DFO suppressecl the expression of epithelial di ffe rentiation markers, such as involucrin, t ransglutaminase II. CK6. and CK19. ancl this suppression was blockecl by p38 ancl ERK MAP kinase ll1hlbltors The oral premalignant(IHOK) ancl malignant cell s(I-lN4) showed differential responses to DFO with respect to the expression of cel l cycle regulatory proteins. cell growth. ancl apoptosis. Coll ectively. the current study reveals that p38 MAP kinase plays an ill1 portant role in iron chela tor-mecliatecl cel l cleath and in the suppression of differentiation of oral premalignantandmalignanLcell s.by activating a c10wnstream apoptotic cascade that executes the ceIl c1eath pathway

Interlellkin • 8(IL-8) is an important cytokine involved in tllmor growth and angiogenesis in a variety of malig nancies. bllt the regll lation of IL-8 in 01 외 cancer cells are llnderstood . We invesLigated whether mi togen-activated protein kinases pathway is involved in iron chelator-mediated lL-8 produdion in inunortalized and malignant oral keratinocytes. In this study we examined the role of p38 and extracellular signal- reglllated kinase• 1/2 in the expression of [L-8 by DFO. Incllbation of IHOK and HN12 cel ls with DF'O increased the expression of 11-8 mRNA. as well as the release of IL-8 protein. The signal transdllction study revealed that both p38 and ERK1/2 were significantly activated in response to DFO. Accord ingly. the selective inhibitors for both kinases‘ eit her a lone or combination. abolished DFO- induced lL-8 secretion. indicating an importance of MAP kinase pathway. Interestingly. however‘ inhibition of the p38 and ERK pathway more attenuated IL-8 secretion in IHOK than in HN12 cells. DFO induced NF-kB activation , suggesting a NF-kB- dependent mechanism in DFO- induced IL-8 production. In addition, p38 and ERK inhi bition resulted in the accelerated degradation of lL-8 mRNA, suggesting that in IHOK and HN12 cells, p38 and ERK cunLr iullLe Lo DFO imluced IL-8 secretion by IHOK and HN12 cells via a posttranscriptional mechanism that involves stabilization 01' the IL-8 transcript. Finally. we investigatecl llsing specific inhibitors : 8NP and G8NO for NO c1onor. PDTC for potent inhibitor of NF-kB. Cycloheximide for inhibition of de novo protein synthesis. We observecl 8NP ancl PD1'C clepenclent IL-8 gene incluction in IHOK cell s. but not in HN12 cells used specific inhibitors‘ Collectively. these results demonstrate that‘ targeting MAP kinase ancl NF-kB pathway may be a potentiaI approacb to controlling the angiogenes is ancl growth 이 human oral cancers

뻐ny studìes have shown the anti-proli ferative effects of irondeprivation on cancer cell s‘ but the effects 01' iron-chelators on oral cancer have not been clearly elucidated , To investigate the effects of an iron chelato r, desferrioxamine( 01"O).on the growth of ilIllTIortali zed human o1'al ke ratinocytes(IHOK), primary oraJ cancer cel ls(HN4)‘ metastatic oral cancer cell s(HN12) , and human skin keratìnocytes(HaCaT) in the MTr assay, three-dimensional(3D) raft cul tmes, Western blott ing, cell cycJ e analysis‘ nuclear staining‘ and cytochrome c expression for apoptosis s ig naling pathway were used OFO inhibited the growth of immortalized IHOK and HaCaT and mal ignant HN4 and HN12 keratinocytes in a time- and dose-dependent manner according to the MTT assay, The 3D organotypic cu l tu re also revealed that OF'O-treated cells showed less epithelial maturation, less surface keratinizati on‘ and de creased epithelia l thickness, The major mechanìsm of growth inhìbition with the micromolar 0 1"0 treatment was by the induction of apoptosis‘ which was supported by nuclear OAPI staining, ONA fragmentation analysis, and J10w cytometric analysis for sub-Gl phase ar rest and Annexin V-1"ITC stainìng, Furthermore‘ Bax expression in creased together with p53 and p21WAF1!CIPl, whìle the Bcl-2 expression decreased in the immortalized and malig nant keratinocytes treated with 01"0 , Time-dependent cytochrome c from mitochondria was observed in D1"O-treated [l-IOK and 0 1'머 cancel‘ ceJJ s, and was accompanied by the activation of caspase-3 in IHOK cells. These resu lts demonstrate that 0 1"0 has growth inhibitory effects on immortalized and malignant oral keratinocyLes Lhrough the induction of apoptosis and suggest that further evaluation of OFO as a potcntial thcrapcutic agent for human oral precancerous and cancerous lesions is warranted

We studi ed the difTerential elTect of vitamins A, C, U. and E on normal human 이 al keratinocyte(NHOK) , HPV-16 E6E7 immor talized human oral keratinocyte(1HOK) , Oncogene transfected HPV-16 immortalized ce1ls(OTOK) , and two ol'al sq ua mous cell line(HNSCC30‘ HNSCC31) according to carcinogenesis stage. The vitamin effect was evaluated by morphology. ce ll viabi lity. a nd orgnaotypic culture Vitamin A has a greater negative effect on growth for all NHOK IHOK HNSCC. es pec ially N-Ras t rans fected IHOK, Vitamin D & E revealed no significant cell activity on NHOK lHOK, ad OTOK Vitamin C was found increased cell viability to IHOK and OTOK 1n primary oral squmaous cell ca rcino ma (HN30 ). vitam in 0 and C showing increased cell growth , but vitamin E showing no effect 1n metastatic oral squamous cell ca rcinoma(HN31), vitamin C has prol iferative effect , but vitamin 0 & E has anti-proliferative effect Vitamin A t reated normal a nd ma lignant ce1ls by organotypic cu lt ure. showed reduction of epithelial layer and in vasion to connective tissue. , especia lly in 1HOK & oncogene-transfected 1HOK, 1n conclusion. three-dimensional culture sys tem may be useful as a model to acess the efficiency of agents such as a1l trans retinoic acid can preventing progression of these premaligant lesion to maligant oral carcinoma(ch emopreventive agent) .

Studies to evaluate distribution of markers in normal keratinocyte and their immortalized keratinocyte are appropriate to evaluate the normal and preneoplastic lesion of oral cancers as biochemical and cytochemical changes associate with tumorigenesis being not completely understood. Complementary DNA microarray containing 6000 sequence -verified cDNA elements was used to systematically characterize the variation in gene expression patterns of NHOK cells vs. immortalized keratinocyte by HPV16 E6-E7(IHOK). Examination of gene expression that is 85 clones cDNAs exhibits greater than 2 fold overexpression in NHOK probes relative to IHOK probe, 147 cDNAs reveal greater 2 fold overexpression in IHOK relative to NHOK probe.The high similarity in gene expression (96.5%) between IHOK and NHOK cells suggests that only an additional 232/6720 (3.5%) of the genome is differentially gene activated during HPV16 immoratlized keratinocyte growth and differentiation. Examination of gene expression that differs between NHOK and IHOK cellsapprear to be related to : cell adhesion & recognition, cell cycle regulator, apoptosis, transciption factors, growth factors and therir receptors, cytoskeletal and extracellular matrix proteins, signal transduction modulators and effectors, and miscellaneous. The gene expression of cell recognition factor such as endothelin 1, collagen IV, fibronectin, and SPR1 in IHOK were upregulated. Distinct or duplicated cDNA clones representing the same gene were typically clustered in adjacent rows in the clustered gene map. Therefore the differentially expressed and identified genes should be informative in studying oral epithelial cell carcinogenesis and such studies should foster the research of molecular markers allowing to assess the phenotypeof malignant epithelial tumor.