The black-veined white, Aporia crataegi (Lepidoptera: Papilionoidea), is nearly extinct in South Korea, although substantial numbers of dried specimens are available. One of the common practices for such species is to launch re-introduction program after proper amount of genetic information are analyzed from donor and donee populations. In this study, we sequenced complete mitochondrial genome (mitogenome) of A. crataegi to design species-specific primers for subsequent population works and to further understand the mitogenome evolution in lepodiopteran Papilionoidea. The 15,140-bp long A. crataegi mitogenome that has typical sets of 37 genes is smallest among true butterfly species with overall slightly smaller size in genes and regions throughout the genome. Arrangement of the genome is identical to those of other lepidopteran mitogenomes, in which tRNA cluster located between the A+T-rich region and ND2 gene is translocated into tRNAMet, tRNAIle, and tRNAGln from ancestral arrangement, tRNAIle, and tRNAGln, tRNAMet. The A/T content of the genome at 81.3% is the highest in Pieridae, but lower than that of lycaenid species (81.7% ~ 82.7%) The high A/T content in the genome is also reflected in codon usage, accounting for 41.69% of A/T-composed codons (TTA, ATT, TTT, and ATA). Unlikely the diversified or modified usage of anticodon for tRNASer(AGN) the species of Pieridae including A. crataegi all unanimously have GCT that has been hypothesized as ancestral for Lepidoptera. A total of 111 bp of non-coding sequences are dispersed in 13 regions, ranging in size from 1–49 bp. Among them relatively longer ones (≥ 16 bp) all have relatively higher sequence identity to other regions of the genome, suggesting partial duplication of the sequences during A. crataegi evolution. As has been reported in some species of Lepidoptera, the A. crataegi A+T-region also has typically found conserved sequences (e.g., poly-T stretch, ATAGA motif, ATTTA element, microsatellite-like A/T sequence, and poly-A stretch) and one tRNA-like sequence, and this feature was commonly found in true butterfly species.

The phylogenetic relationships among the Nymphalidae (Lepidoptera: Papilionoidea) have been controversial in several perspective. The present study sequenced a total of ~ 3,500 bp from cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-1 alpha (EF-1α) in 80 nymphalid species belonging to seven subfamilies (Linmenitidinae, Heliconiinae, Nymphalinae, Apaturinae, Libytheinae, Satyrinae, and Danainae), along with those of six lycaenid species as outgroups. Phylogenetic analyses via Bayesian Inference (BI) and Maximum Likelihood (ML) algorithms concordantly supported the subfamilial relationships of (((((Linmenitidinae + Heliconiinae) + (Nymphalinae + Apaturinae)) + Libytheinae) + Satyrinae) + Danainae), with high nodal support for monophyletic subfamilies and tribes. This result is largely consistent with a previous study performed with a substantially large sequence information and morphological characters, except for the position of Libytheinae that has previously been placed as the sister to all reminder of Nymphalidae.

운간초의 분화 품질을 향상시키기 위하여 ‘Kumoma’ 와 ‘Kumoma-Gusa’ 두 품종의 생장과 개화에 미치는 영향을 구명하고자 식물생장억제제 (PGRs) 4종을 처리 하였다. 식물생장억제제 종류와 처리농도는 paclobutrazol (10, 20, 40, 80 mg·L-1), flurprimidol (5, 10, 20, 40mg·L-1), daminozide (500, 1000, 2000, 4000mg·L-1), chlormequat (50, 100, 200, 400 mg·L-1)이고 엽면살포 와 토양관주 방법으로 처리하였다. Paclobutrazol 40 mg·L-1 처리는 운간초의 두 품종에서 초장과 화경 장을 줄이는데 효과적이었다. ‘Kumoma’ 품종에서, paclobutrazol 40 mg·L-1 엽면살포와 토양관주 처리방 법은 초장을 각각 12.6, 12.5 cm로, 화경장은 3.4, 3.3 cm로 줄일 수 있었다. ‘Kumoma-Gusa‘ 품종에서 는 paclobutrazol 토양관주는 처리 농도가 증가함에 따라 초장은 13.2-10.7 cm, 화경장은 3.9-2.0 cm까지 줄일 수 있었지만 꽃의 수는 대조구와 비교했을 때 현 저하게 감소하여서 20개까지 감소하였다. Flurprimidol 20mg·L-1 스프레이와 토양관주 처리는 두 품종 모두 초장과 화경장을 줄일 수 있었다. ‘Kumoma’ 품종에 서 flurprimidol 10 mg·L-1 스프레이 또는 토양관주 처리는 개화 품질을 향상시킬 수 있었고 대조구와 비 교해 꽃의 수는 44.7개 이상, 출하 일수는 5일 정도 빨리 출하 할 수 있었다. ‘Kumoma-Gusa‘ 품종에서 PGRs처리에 의한 초장 과 화경장 억제효과가 가장 큰 것은 paclobutrazol 80mg·L-1과 flurprimidol 40mg·L-1 이었고 paclobutrazol 과 flurprimidol 스프레이와 토양관주 처리에서 각각, 초장은 10.7, 9.9 cm, 화경장은 2.0, 1.5 cm 로 생장 억제효과가 가장 큰 것을 알 수 있었다. 그러나 flurprimidol 40 mg·L-1 토양관주 처리에서 출하일수는 농도에 따라 3-13일 정도 대조구보다 늦어졌고 꽃의 수는 15.6개로 가장 적은 것을 관찰 할 수 있었다. 두 품종 모두에서 daminozide 처리는 초장과 화경장 이 줄어들지 않았지만 ‘Kumoma’ 품종에서는 대조구 와 비교했을 때 꽃의 수가 증가하였다. Chlormequat 처리는 paclobutrazol 과 flurprimidol 처리와 비교했 을 때 화경장 감소효과는 작았고 처리 식물의 잎은 다른 처리에 비해 연한 녹색을 보여주었다. 두 품종 모두에서 chlormequat 과 daminozide 처리는 생육과 개화 품질에 효과적으로 영향을 주지는 않았지만 꽃의 수는 모든 처리농도에서 41개 이상을 보여주었고, ‘Kumoma-Gusa’ 품종에 chlormequat 토양관주 처리는 최대 꽃의 수 63개를 볼 수 있었다. 이번 결과들은 paclobutrazol 과 flurprimidol 처리가 운간초 분화의 품질 향상을 위한 화경장의 생육을 조 절하는 생장억제제로서 사용 될 수 있음을 보여 주었 고, ‘Kumoma’ 품종에서는 paclobutrazol 40 mg·L-1 스프레이는 생육특성을, flurprimidol 10 mg·L-1 토양관 주 방법은 개화관련 특성의 품질을 향상시키는 생장억 제제의 농도로 추천 될 수 있고, ‘Kumoma-Gusa’ 품종 에서는 flurprimidol 20mg·L-1 스프레이와 flurprimidol 5mg·L-1 토양관주 방법이 운간초 분화의 생육특성과 개 화품질을 향상시키는 생장억제제의 농도로 추천 될 수 있다.

The Samia cynthia ricini (Lepidoptera: Saturniidae) is a commercial silk-producing insect belonging to an insect family Saturniidae in Bombycoidea. The species that has presumably been originated in India, is distributed in India, China, and Japan. Unlikely domestic silkworm the prime host plant for the species is a castor-oil plant (Ricinus communis in Euphorbiaceae). Recently, the eri-silkworm also is reared in Korea and is expected to be utilized for a diverse purpose. In this report, we present the complete mitochondrial genome of the species with the emphasis of a few major characteristics. The 15,384-bp long S. cynthia ricini (Lepidoptera: Saturniidae) mitochondrial genome was amplified into three long overlapping fragments (from COI ~ ND4, ND5 ~ lrRNA, and lrRNA ~ COI) and subsequent several short fragments using the long fragments as temperate. The primers for both long and short fragments were designed solely for lepidopteran genomes, without any species-specific primers. As a usual the genome is composed of 37 genes: 13 protein-coding genes (PCGs), two rRNA genes, and 22 tRNA genes, and one large non-coding region termed the A+T-rich region. Arrangement of the genome is identical to those of other lepidopteran mitochondrial genome, but this differs from the common arrangement found in a diverse insect order, by the movement of tRNAMet to a position 5’- up stream of tRNAIle. Unlikely previous report on the start codon for COI gene in Lepidoptera S. cynthia ricini COI gene starts with typical ATT codon located between tRNATyr and the beginning region of COI gene. The 22 tRNAs that are interspersed throughout the mitogenome ranged in length from 62 to 71 bp. All tRNAs but tRNASer(AGN) were shown to be folded into the expected cloverleaf secondary structures. More detailed structural and phylogenetic analyses among Bombycidae and Saturniidae in connection with other families in the Bombycoidea will be performed soon

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation of the species we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) collected from seven Korean localities. A total of 18 haplotypes (BARCA01 ~ BARCA18), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 17 sequence types (ITS2CA01 ~ ITS2CA17), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in mitochondrial gene sequence. Phylogenetically, the mitochondrial DNA has shown several haplotypes formed independent groups with substantially high node support (≥ 90%), whereas no such grouping was evidenced for ITS2, indicating different behaviors of the two molecules. Such difference may reflect a diverse dynamics of the species such as biogeographic history, mating behaviors, and also possibly different mode of inheritance of the two molecules, but requires further scrutinized examination of the dataset. In terms of population genetic perspective, overall no population subdivision was detected from both molecules, except for locality 7 (Eocheong islet) from mitochondrial DNA. As more scrutinized analysis is performed, further fruitful inference on the geographic contour of the species might be available.

We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.

The principal objective of this study was to assess the antioxidative activities of Petasites japonicus against oxidative stress in bovine brain tissue . Petasites japonicus is found with a relatively widespread distribution, and is cultivated as a culinary vegetable in Korea. Petasites japonicus samples were dried either by freeze-drying or by hot air-convection drying (80℃), then evaluated for their antioxidative activity by measuring 1-dipheny-1，2-picrylhydrazyl (DPPH) radical scavenging, and by measuring thiobarbituric acid-reactive substances (TBARS) in brain homogenates subjected to Fe2+ -mediated lipids with or without the addition of botanical extract. Hot air convection-drying resulted in a slight increase in the extraction yie1d as compared with freeze-drying. However, total phenol and flavonoid contents in freeze-dried Petasites japonicas were significantly higher than those of hot air convection-drying. Freeze-drying increased the free radical scavenging activity of Petasites japonicas, leaves, and stems by 52.6, 28.6, and 248.0%, as compared with hot air convection-drying. Additionally, the IC50 values measured by TBARS in hot air convection-dried Petasites japonicas， leaves, and stems were increased by 36.0, 31.6, and 15.9%, as compared to those of freeze-drying. Although antioxidative activity was reduced slightly by heat processing in Petasites japonicas, freeze-drying for each portion of Petasites japonicus was the most appropriate for use as a functional food and pharmaceutical material.

Physicochemical qualities and consumer acceptability of chocolate layer cake were studied with varied levels of rosemary powder at 0, 0.2, 0.4 and 0.6%. The ash content of the cake increased from 2.30 to 3.10%, as the amount of rosemary powder increased from 0 to 0.6%, and the carbohydrate content of the cake decreased as the addition of rosemary powder increased. There were no significant differences in moisture contents and pH values among the samples and the pH values of all samples were within the typical pH range of 7.5-8.0 for chocolate， layer cakes. Water loss from the control cake was greater than that from the cakes with rosemary powder supporting the suggestion that the addition of rosemary powder to the chocolate layer cake could increase moisture retention of the cake. Consumer acceptability of all the samples showed higher prefierences of more than 7 points. Rosemary aroma, mint flavor and after taste were highly positively corrεlated with the fat content. Fat and ash content of the cake, which tended to increase in proportion to the rosemary powder content， were negatively correlated with acceptance of herb flavor, sweet taste, moistness， softness and intensity of softness but positivεly correlated with intensity of herb flavor. With the results above, trials on chocolate layer cake using rosemary powder were successfully performed within the ranges tested.

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

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