Mealworms, Tenebrio molitor (L.) is used as an important animal feed additive for growth promotion and health management, but potentially exposes to fungal infection. In this work, virulence of two species of entomopathogenic fungi against the insect, and the relationship between abiotic features and virulence were investigated. Secondly our consideration was also given to the effect of chemical fungicides on conidial germination for risk control. Between Beauveria bassiana (Bb) and Metarhizium roberstii (Mr) (previously M. anisopliae), Bb isolates had much higher virulence (~100% mortality in 3~4 days after the treatment), rather than Mr isolates in laboratory assays. Next, fungus-treated mealworms were kept at wheat bran at 20, 25, 30 and 35℃ with 3, 6, 9 times of water spray to the feeds for set-up of different humidity conditions. Inoculation of fungi to mealworms was conducted by fungal spray and feeding methods, which resulted in higher virulence in feeding method. In the feeding method, all temperature treatments except 35℃ showed high virulence against mealworms, but any significant relationship between virulence and humidity was not observed. In the chemical fungicide screening, fluazinam (CAS No. 79622-59-6) and mancozeb (8018-01-7) significantly inhibited the germination of Bb and Mr conidia. This work suggest that contamination of wheat bran with fungal pathogens, particularly B. bassiana may induce mycosis of mealworms, but introduction of effective fungicides possibly reduce fungal infection.

Entomopathogenic fungus, Beauveria bassiana can be used in integrated crop management and pharmaceutical applications. Recently some efforts have been given to the fungus to improve its biological performances, but low fungal transformation efficiency is one of the limitations. In this work, B. bassiana ERL1170 isolate was used for fungal transformation by restriction enzyme-mediated integration, where pBARKS1-Bbs-cecropinA was linearied using HindIII. The fungal transformation comprised two steps, preparation of competent blastospores and integration of the plasmid into the cells. To prepare competent blastospores, 2-d cultured blastospores were individually treated with 0.1, 0.2, 0.4 and 0.8 M lithium acetate (LiAc). Secondly in the integration step, concentration of LiAc and calcium chloride (CaCl2), and time period of heat shock were investigated as follows: LiAc, 1, 2 and 4 M; CaCl2, 50, 100 and 200 mM; and heat shock at 42℃, 20, 40 and 60 min. Consequently, combination of 0.2 M LiAc in preparing competent blastospores and 2 M LiAc and 200 mM CaCl2 in the second step showed the highest transformation efficiency. This work would be helpful in the fungal transformation of B. bassiana.

This study is intended to examine the motor skill learning and treadmill exercise on motor performance and synaptic plasticity in the cerebellar injured rats by harmaline. Experiment groups were divided into four groups and assigned 15 rats to each group. GroupⅠ was a normal control group(induced by saline); GroupⅡ was a experimental control group(cerebellar injured by harmaline); GroupⅢ was a group of motor skill learning after cerebellar injured by harmaline; GroupⅣ was a group of treadmill exercise after cerebellar injured by harmaline. In motor performance test, the outcome of groupⅡ was significantly lower than the groupⅢ, Ⅳ(especially groupⅢ)(p<.001). In histological finding, the experimental groups were destroy of dendrities and nucleus of cerebellar neurons. GroupⅢ, Ⅳ were decreased in degeneration of cerebellar neurons(especially groupⅢ). In immunohistochemistric response of synaptophysin in cerebellar cortex, experimental groups were decreased than groupⅠ. GroupⅢ's expression of synaptophysin was more increased than groupⅡ, Ⅳ. In electron microscopy finding, the experimental groups were degenerated of Purkinje cell. These result suggest that improved motor performance by motor skill learning after harmaline induced is associated with dynamically altered expression of synaptophysin in cerebellar cortex and that is related with synaptic plasticity.

Sulfur is commonly used in Asia as an herbal medicine to treat inflammation and cancer , a nd potent chemo preventi ve effects have been demons tra ted in various in vivo and in vitromodels for s ul fur-containing compounds found in natura l1y occurring product s. Here, we 1'eport the growth inhibitory and apoptosis-related effects of a n ewly developedhigh-puri ty edible sulfur(ES) on immo1'tali zed human o1'al ke1'atinocytes(IHOKs) and on oral cancer cells representing two stages of oral cancer (HN4‘ HN12) based on an 3-(4,5-Dimethylthiazol-2-yl) -2.5- diphe n yltetrazolium bromide(MTI) assay, Western blotting, cell cycle analysis, and nuclear staining. The puri ty of the ES used in thi s study was ve1'ified by high performance liquid chromatog1'aphy (HPLC) , ami no acid analysis and energy dispersive spectroscopy(EDS). ES inhibited the prolife1'ation of imrnortalized and ma lig nant o1'al kerati nocytes in a dose- and time-dependent manne1' FITC-Annex.in V staining, DNA fragmentation t esting. and Hoechst 33258 s taining revealed that ES inhibits cell growth via apoptosis. ES bl ocked cell-cycle prog1'ession at t he sub-Gl phase‘ wi th decreased expression of cyclins Dl, D2‘ and E, and their activating partn ers cdk2‘ cdk4‘ and cdkfì, and a concomitant induction of p53 and p21/WAF1. Furthe1'more, ES treatment in creased the cytosolic level of cytochrome c and resulted in caspase- 3 activation‘ and thi s effect was co1'1'elated with Bax up-regulation and Bcl-2 down-1'egulation Taken together‘ these data suggest that ES is a potential chemopreventive and chemotherapeut ic agent fo r oral ca ncer

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

This study was to taken to demonstrate the effects of exogenous nitric oxide(NO) on hu rnan pu lp cell s ‘ In volvement of cyclic 3’, 5' -monophosphate(cGMP) in p버 paJ protection induced by herne oxygenase-l (J-lO-l) against NO-induced cytotoxicity , By use of Western blotting and cell viabi lity assay, we have examined the cytotoxicity and J-lO-l induction in pulp cells that were treated with NO donor ‘ S-nitroso-N-acetyl-D, L-penici 1 lamine(SNAP) , We have assessed wheathel' HQ--l contributes the cytoprotective effect against the cytotoxicity caused by NO, and inves tigated the l'elationship between HO-l and cGMP in the s ignaling pathway, SNAP decreased cell via bility but in creased HO-l expl'ession in a concentl'ation- and time一dependent manner in hurnan pu lp cells NO-induced cyto toxicity was inhibited in the presence of the hemin(inducer of HO-l) , whel'eas was en hanced in the pl'esence zinc protoporphyrin IX(ZnPP IX, HO-l inhibitor), thus Lhe NO-induced cytoLoxicity was cOl'related with HO- l expression. R‘ etreatment with a rnemhrane-permeable cGMP analog, 8-bromo-cGMP, restored cell death and enhanced the HO-l protein expression induced by SNAP, ln contrast‘ inhibition of guanylate cyclase by lI-l -[1,2,4] ox adiazole[ 4,3 口]quinoxalin-l-one(ODQ) pretreated pulp cells to 1 mM SNAP resulting in marked cytotoxicity , These findings , demonstrating a link between J-lO-l, regulated thl'ough the cGMP system and NO-induced cytotox.icity in huma띠 p버 p ceJls , suggesti ng a protective 1'ole of HO-l in pulp infl ammatory disease

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

본 연구는 일반인을 대상으로 시대성이 반영된 각종 잠재적 위험요소와 원자력(방사선)발전의 위험성 대 한 인식을 분석하였다. 설문 대상은 다양한 계층으로 하고 그 중에 총 293부를 분석하였다. 그 결과, 잠재 적 위험요소 중에서 화재에 대한 위험도 인식이 높게 나타났으며, 다음으로 방사선 테러와 핵(원자력)에너 지의 위험도를 다른 위험요소들에 비해 비교적 높게 인식하였다. 연령별, 학력별, 정치 이념 성향에 따른 분석에서 원자력발전의 필요성과 위험성, 안전성에 대해서 상반된 결과를 나타내었다. 정치이념의 성향에 따른 잠재적 위험요소와 원자력발전에 대한 인식은 보수적 이념집단에서 긍정적인 인식을, 진보적 집단에 서는 부정적인 인식을 하고 있는 것으로 나타났다. 즉, 정치 이념의 성향에 따라 원자력발전 인식 분석에서 통계적 차이를 보였다. 따라서 원자력(방사선)발전 정책 방향 설정과 방사선 이용 관련 산업에 있어서 전문 가 의견과 일반인의 다양한 의견을 반영해서 결정되어져야 할 것으로 판단되고, 일반인도 사실을 바탕으로 객관적이고 과학적인 근거에 입각해 각종 잠재적 위험과 원자력(방사선)에 대해 막연한 불안감을 가지지 말고 유연한 대처를 할 필요가 있다고 판단된다.

Background : Ixeris genus has been used in traditional medicines as stomachics, sedatives, and diuretics. Ixeris dentata var albiflora is a kind of perennial herbaceous plant and one of the plants of the genus Ixeris (Asteraceae). It is well-known for edible wild vegetable in Korea, China, Japan, and Mongolia. Specially, Korean has its root and young leaf with appetizing vegetable due to bitter taste. Methods and Results : We isolated 8 genes that are involved in carotenoid biosynthesis using the Illumina/Solexa HiSeq2000 platform. In this study, a full-length cDNA clone encoding phytoene synthase (IdPSY), phytoene desaturase (IdPDS), ξ-carotene desaturase (IdZDS), lycopene β-cyclase (IdLCYB), and zeaxanthin epoxidase (IdZEP) and partial-length cDNA clones encoding lycopene ε-cyclase (IdLCYE), ε-ring carotene hydroxylase (IdCHXE), and β-ring carotene hydroxylase (IdCHXB2) were identified in I. dentata. The theoretical molecular weight (MW) and isoelectric point values of 8 genes were investigated. Sequence analyses revealed that these proteins shared high identity and conserved domains with their orthologous genes. IdPSY, IdPDS, IdZDS, IdLCYB, IdCHXB2, and IdZEP were constitutively expressed in the roots, stems, leaves, and flowers of I. dentata. Conclusion : Our study on the biosynthesis of carotenoids in I. dentata will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of I. dentata.

Background : Platycodon grandiflorum has been used as food material and a traditional medicine in Korea. In order to develop an efficient in vitro micropropagation technique for a rare plant species and conservation for inbred line of plant breeding. Methods and Results : Plant regeneration via organogenesis and somatic embryogenesis was investigated in Platycodon grandiflorum. Leaf, stem, root tissues of 7-day-old seedlings were cultured on 1/2MS medium containing various concentration (0, 0.5, 1 and 2 ㎎/L) of IBA, BA and NAA. The results showed that 1/2MS medium supplemented with BA+NAA 2.0 ㎎/L yielded the highest callus formation ratio of 73.5%. When various tissues (leaf, stem, root) were tested on 1/2MS medium containing BA 2.0 ㎎/L+ NAA 2.0 ㎎/L for somatic embryogenesis, the optimum tissue for embryogenic shoot induction was stem tissue. Callus were cultured on MS medium containing various concentration (0, 0.5 and 1 ㎎/L) of BA and NAA. The best rooting rate was achieved by three different medium (B5, 1/2MS and MS) and 1/2MS medium cultured the highest rooting ratio (82%). Conclusion : This study suggested that above micropropagation techniques can be used for rapid multiplication as well as in vitro or in vivo conservation of the Platycodon species.

Background : This study was investigated the effect of organic fertilizer application and stem training methods on the growth and yield of Cynanchum willfordii. Methods and Results : Traditional species, C. willfordii conducted the test to the field in Chungbuk ARES, Korea. Two methods, non-supporting (custom) and I-shape supporting was used for stem training method. Planting spacing was adjusted to 30 (interrow spacing) x 20 cm (intrarow spacing) and other key management is the followed the cultivation manual for standard medicinal crops. Organic fertilizer application is conducted in mid-March before planting of C. willfordii. Registered organic agricultural materials such as fungal cultures (CC), mixed organic materials (MO), fermented and mixed expeller cake (EC), and fermented fowl dropping (FD) was used. Application rate is based on the nitrogen application rate after soil testing. Plant height was both good in all at I-shape supporting (266.7 ㎝) and all in non-supporting (160.8 ㎝); however, stem diameter was more thicker in custom (4.6 ㎜) of I-shape supporting and EC (4.8 mm) of non-supporting. Number of branched stem were good at EC from I-shape supporting and non-supporting, as well as leaf growth. SPAD value was evaluated in MO (52.7) of I-shape supporting and EC (52.3) of non-supporting. Number of branched root per organic fertilizer were showed 7.0 at MO from I-shape supporting, but 7.3 at FD from non-supporting. While length of main root were 27.0 cm in MO of I-shape supporting and 31.3 ㎝ in FD of non-supporting. Root weight also more heavier in EC (66.3 g) of I-shape supporting and FD (53.0 g) of non-supporting. When applied organic fertilizer, total density of soil microorganisms were changed into 38.4 ppm in custom; however, it more plentiful of 90.7 ppm in MO, it showed good effect on the replication of soil microbiota. Conclusion : From the investigated results, MO of I-shape supporting was good at overall growth, including good tendency of roots growth.

Background : The effect of ridge-up bed cultivation and stem training method on the plant growth and yield of Cynanachum willfordii was investigated. Methods and Results : Domestic variety of Cynanachum willfordii was tested at Chungbuk ARES under the following conditions. Nonsupporting (custom) and I-form supporting was used for stem training method, and ridge height was set as 30 (custom) and 60 cm (high ridge). Planting spacing was adjusted to 30 (interrow spacing) × 20 cm (intrarow spacing), and other major management was followed the method of standard cultivation for medicinal crop. Investigated result from leaf characteristics, leaf length was longer in high ridge cultivation (HRC) as 11.1 cm than custom cultivation (CC) as 10.6 cm. Leaf width is proved to be 12.8 (HRC) > 11.2 cm (CC). Leaf number is proved to be 294 (HRC) > 254/plant (CC), with higher number of 44/plant at HRC and weight/10 leaves were more heavy at HRC (4.9 g) than that of CC (2.6 g). It was more fruit setting at HRC over 15/plant. According to the stem training method and ridge height from nonsupporting cultivation, main root was 4.0 (CC), higher than that of HRC over 0.5/plant. However, root length was more longer in HRC (28.6) than that of CC (25.0 cm). Main root diameter was also showed more thicker pattern in HRC. From staking cultivation, root number of HRC was 7/plant, it was recorded more 3/plant than that of CC, and it was also same pattern in main root length and root diameter. By the standard of commercial root, yield of living roots in nonsupporting cultivation were 59.0g/plant (HRC), it was more heavy over 10.4g/plant than that of CC. In staking cultivation, HRC were recorded as 74.2g/plant, more heavy 6.9g/plant than that of CC. Yield from I-form support stem training and ridge-up bed cultivation of HRC was higher approxmiately 52.7% than that of CC. Conclusion : Overall growth by high ridge cultivation of I-form support was good and yield of HRC was also increased over 52.7% than that of CC.

May-Thurner syndrome is caused by blockade of local venous flow due to local vascular intimal proliferation, caused by repeated pulsatile compression of the iliac or iliofemoral vein between the iliac artery and the lumbar spine. In this case, we confirmed May-Thurner syndrome using lower extremity computed tomographic angiography and venography. However, on venography, it was impossible to distinguish the left iliac vein from the collateral vein; a thrombus was also seen, although some of the thrombus was not seen clearly. These problems were overcome with use of intravascular ultrasound. We report on intravascular ultrasound guided treatment of May-Thurner syndrome.

May-Thurner syndrome is associated with deep vein thrombosis resulting from chronic compression of the iliac vein against the lumbar vertebrae caused by the overlying common iliac artery. Stent insertion into the compressed lesion is used in treatment of May-Thurner syndrome. Various complications can occur during angioplasty while using a stent. Among these complications, shrinkage of the vein below the stent, a rare complication, was observed in our hospital during treatment of a patient with May-Thurner syndrome. Different complications can occur when venous angioplasty is performed, unlike that when arterial angioplasty is performed.