Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA2+ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

Determining the infection history of living organisms is essential for understanding the evolution of infection agents with their host, particularly for key aspects such as immunity. Viruses, which can spread between individuals and often cause disease, have been widely examined. The increasing availability of fish genome sequences has provided specific insights into the diversity and host distribution of retroviruses in fish. The shortspine spurdog (Squalus mitsukurii ) is an important elasmobranch species; this medium-sized dogfish typically lives at depths of 100~500 m. However, the retroviral envelope polyprotein in dogfish has not been examined. Thus, the aim of the present study was to identify and analyze the retroviral envelope polyprotein in various tissues of dogfish. The 1334-base pair full-length novel cDNA of dogfish envelope polyprotein (dEnv) was obtained by 3' and 5'-rapid amplification of cDNA end analysis from S. mitsukurii. The open reading frame showed a complete coding sequence of 815 base pairs with a deduced peptide sequence of 183 amino acids that exhibited 34~50% identity with other fish and bird species. It was also expressed according to reverse transcription and real-time polymerase chain reaction in the kidney, liver, intestine, and lung, but not in the gill. This distribution can be assessed by identifying and analyzing endogenous retroviruses in fish, which consists of three main genes: gag, pol and env. Dogfish envelope polyprotein sequence is likely important in evolution and induces rearrangements, altering the regulatory and coding sequences. This is the first report of the identification and molecular characterization of retroviral envelope polyprotein in various tissues of S. mitsukurii.

Carbonic anhydrase is essential for the cellular transportation of hydrogen and bicarbonate ions and plays a key role in a wide variety of physiological processes. Rainbow trout, Oncorhynchus mykiss is an important freshwater fish in aquaculture industry and is known to be one of the most susceptible species to environmental contamination. In this study, carbonic anhydrase was detected in the kidney and intestine of rainbow trout. Carbonic anhydrase was isolated from cytosolic proteins and identified by using SDS-PAGE, isoelectric focusing, and immunohistochemical methods. A specific protein band with molecular weight of 30 kDa and pI of 7.0 was detected by Western blotting. The immunohistochemical results showed that carbonic anhydrase was located at various cells in the kidney and intestine of rainbow trout.

Entomopathogenic fungi infect some arthropod pest and have been used for biological control. Some entomopathogenic fungi have high potential in insect pest management worldwide, and most of researches were given to Hypocreales order of Ascomycota, but little in Zygomycota, such as Entomophthorales in our country. We have identified some species belonging to Entomophthorales and investigated cultural features and ecology of Entomophaga aulicae. E. aulicae in sweet potato fields, where Aedia leucomelas was a dominant pest, had caused epizootics from 2002–2005. E. aulicae-infected A. leucomelas larva were mostly found from August to October and its occurrence was significantly related to the precipitation. E. aulicae were mainly observed in dead A. leucomelas larval populations (infection rate = 41.3% in 2002), rather than other lepidopteran pests. E. aulicae was morphologically and genetically identified and its virulence was characterized in laboratory conditions. Additionally, pest infection by Zoophthora radicans, Neozygites floridana and some Entomophthorales have been identified. On the basis with this information, we need research to predict the prevalence and to develop biological control agent using Entomophthorales including E. aulicae that contribute to regulation of host populations.

The Hypsizigus marmoreus has been used as medicinal and food source in worldwide. Viridans Streptococci, which include Streptococcus sanguinis and Streptococcus gordonii, are pioneer oral bacteria that initiate dental plaque formation. This study was to investigate the antimicrobial activity of Hypsizigus marmoreus Extracts. Antimicrobial activity of extracts were tested against pathogenic bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus sanguinis and Streptococcus gordonii) by paper disc methods. The Hypsizigus marmoreus Extracts were extracted with 80% methanol. The 80% methanol extracts of Hypsizigus marmoreus Extracts showed antimicrobial activity against S. sanguinis and S. gordonii. The 80% methanol extract of Hypsizigus marmoreus Extracts were fractioned by hexane, chloroform, ethylacetate and buthanol. The ethylacetate fraction of Hypsizigus marmoreus Extracts showed strong antimicrobial activity against S. sanguinis and S. gordonii with minimum inhibitory concentration (MIC) range of 0.8-1 mg/ml. Especially, the 80% methanol extract and ethylacetate fraction of Hypsizigus marmoreus Extracts were inhibited the biofilm formation of S. sanguinis and S. gordonii at a concentration of 1.0 mg/ml. These results suggest that Hypsizigus marmoreus extracts can be developed as a potent antimicrobial agent.

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

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

Research Institute (HARI), NTCS, RDA, in 2004. This cultivar has a short grain shape and about 123 days growth duration from trans-planting to harvesting under the reclaimed saline area of the south-western and the mid-western coastal plain and Honam plai