Putative cadherin genes, which are a receptor of the Bacillus thuringinesis toxins, were predicted from a whole genome sequencing data from the diamondback moth, Plutella xylostella. After the sequence and expression analysis, a Bt receptor cadherin gene was selected. The P. xylostella cadherin gene (PxCad1, GenBank Accession no. GU901158.1) encodes 11 cadherin repeats and a transmembrane domain. The PxCad1 gene was expressed in all developmental stage specifically in gut tissue by RT-PCR analysis. Expression of PxCad1 gene was suppressed by feeding of its specific dsRNA PxCad1 in 4th instar larval stage. The suppression of PxCad1 expression did not significantly feeding of its specific dsRNA PxCad1 in 4th instar larval stage. The suppression of PxCad1 expression did not significantly influence on pupal and adult development of P. xylostella. However, the larval treated with dsRNA PeCad1 (150 ng/larva) significantly reduced susceptibility to B. thuringiensis Cry1Ac (4.83 μg/ml). By contrast, the dsRNA PxCad1 -treated larvae did not show any change in susceptibility to B. thuringiensis Cry1Ca (0.24 μg/ml). These results suggest that PxCad1 is a specific receptor of Cry1Ac toxin from B. thuringiensis in P. xylostella.

A nuclear receptor, Met, mediates juvenile hormone (JH) action to control gene expressions associated with metamorphosis in many insects. In this study, we showed that RNA interference (RNAi) of the Met or Kruffel homolog 1 (Kr-h1) induced the precocious metamorphosis of Tribolium castaneum larvae. JH significantly inhibited cellular immune response of T. castaneum hemocyte by suppressing hemocyte-spreading behaviour and nodule formation in response to bacterial injection. However, either RNAi of Met or Kr-h1 expression did not prevent the JH-inhibitory effect on hemocyte behaviors. However, several inhibitors specific to JH membrane action significantly inhibit the JH action hemocytes. These results suggest that JH responsiveness of hemocyte is not mediated by the nuclear receptor.

Tumor necrosis factor alpha (TNFα) is a multifunctional inflammatory cytokine that regulates various cellular and bio-logical processes. Increased levels of TNFα have been im-plicated in a number of human diseases including diabetes and arthritis. Sympathetic nervous system stimulation via the beta2-adrenergic receptor (β2AR) in osteoblasts suppresses osteogenic activity. We previously reported that TNFα up- regulates β2AR expression in murine osteoblastic cells and that this modulation is associated with TNFα inhibition of osteoblast differentiation. In our present study, we explored whether TNFα induces β2AR expression in human osteo-blasts and then identified the downstream signaling path-way. Our results indicated that β2AR expression was increa-sed in Saos-2 and C2C12 cells by TNFα treatment, and that this increase was blocked by the inhibition of NF-κB acti-vation. Chromatin immunoprecipitation and luciferase reporter assay results indicated that NF-κB directly binds to its cog-nate elements on the β2AR promoter and thereby stimulates β2AR expression. These findings suggest that the activation of TNFα signaling in osteoblastic cells leads to an upregu-lation of β2AR and also that TNFα induces β2AR exp-ression in an NF-κB-dependent manner.

New therapeutic measure are needed to improve the outcome for patients with oral squamous cell carcinoma(OSCC) because OSCC continues to portend a relatively unfavorable prognosis. Recently RNA interference(RNAi) has emerged as an effective method to target specific genes for silencing. Although overexpression of urokinase-type plasminogen activator receptor(uPAR) has been implicated in progression and metastasis of OSCC, the transfection effect of RNAi- uPAR on OSCC has been rarely reported. The purpose of this study were to examine the efficient and specific inhibition of uPAR mRNA and protein expression by siRNA targeting of uPAR through RT-PCR and immunoslot blotting, and to study cell proliferation activity, adhesion, invasion and migration in vitro compared to the controls. In MTT assay, siRNA-uPAR transfected cells showed about 70-80% cell proliferation compared to OSCC cell lines after 2 days. In adhesion assay, siRNA-uPAR transfected cells showed about 20-30% adhesion activity compared to OSCC cell lines, but similar features to those of BSA coated wells. In migration assay, siRNA-uPAR transfected cells showed about 60% migration activity compared to OSCC cell lines, but higher 3.5 folds to those of BSA coated wells. In invasion assay, siRNA-uPAR transfected cells showed about 55% invasive activity compared to parental cell lines. mRNA expression of siRNA-uPAR transfected cells showed about 10-15 % compared to parental cell lines by RT-PCR. Protein expression of siRNA-uPAR transfected cells showed about 25% compared to parental cell lines by ELISA assay. It suggested that RNAi-uPAR tranfection might be used as a potent and specific therapeutic tool for the treatment of oral squamous cell carcinoma, especially in inhibiting invasion and metastasis.

Insects and animals can recognize surrounding environments by detecting thousands of chemical odorants. Olfaction is a complicated process that begins in the olfactory epithelium with the specific binding of volatile odorant molecules to dedicated olfactory receptors (ORs). OR proteins are encoded by the largest gene superfamily in the mammalian genome. We report here the whole genome analysis of the olfactory receptor genes of S. scrofa using conserved OR gene specific motifs and known OR protein sequences from diverse species. We identified 1,301 OR related sequences from the S. scrofa genome including 1,113 functional OR genes and 188 pseudogenes. OR genes were located in 46 different regions on 16 pig chromosomes. We classified the ORs into 17 families, three Class I and 14 Class II families, and further grouped them into 349 subfamilies. We also identified inter- and intra-chromosomal duplications of OR genes residing on 11 chromosomes. A significant number of pig OR genes (n=212) showed less than 60% amino acid sequence similarity to known OR genes of other species. We also performed a similar analysis on the cattle OR subgenome and identified 1,071 OR related sequences. We show that S. scrofa has one of the largest OR repertoires, suggesting an expansion of OR genes in the swine genome. Considering available information from literature, it seems that OR systems between mammals and insects possess high similarity in their action mechanisms and rapid evolutionary changes due to differences in living environments.

Receptor activator of NF-κB ligand (RANKL) is an essential cytokine for osteoclast differentiation, activation and survival. T lymphocytes such as T₁₇ cells, a subset of T helper cells that produce IL-17, play an important role in rheumatoid arthritic bone resorption by producing inflammatory cytokines and RANKL. It has not yet been clearly elucidated how T cell activation induces RANKL expression. T cell receptor activation induces the activation of nuclear factor of activated T cell (NFAT) and expression of its target genes. In this study, we examined the role of NFAT in T cell activation- induced RANKL expression. EL-4, a murine T lymphocytic cell line, was used. When T cell activation was induced by phorbol 12-myristate 13-acetate (PMA) and ionomycin, RANKL expression increased in a time-dependent manner. In the presence of cyclosporin, an inhibitor of NFAT activation, this PMA/ionomycin‐induced RANKL expression was blocked. Overexpression of either NFATc1 or NFATc3 induced RANKL expression. Chromatin immunoprecipitation results demonstrated that PMA/ionomycin treatment induced the binding of NFATc1 and NFATc3 to the mouse RANKL gene promoter. These results suggest that NFATc1 and NFATc3 mediates T cell receptor activationinduced RANKL expression in T lymphocytes.

The GroEL heat-shock protein from Fusobacterium nucleatum, a periodontopathogen, activates risk factors for atherosclerosis in human microvascular endothelial cells (HMEC-1) and ApoE-/- mice. In this study, we analyzed the signaling pathways by which F. nucleatum GroEL induces the proinflammatory factors in HMEC-1 cells known to be risk factors associated with the development of atherosclerosis and identified the cellular receptor used by GroEL. The MAPK and NF-κB signaling pathways were found to be activated by GroEL to induce the expression of interleukin- 8 (IL-8), monocyte chemoattractant protein 1 (MCP- 1), intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), E-selectin, and tissue factor (TF). These effects were inhibited by a TLR4 knockdown. Our results thus indicate that TLR4 is a key receptor that mediates the interaction of F. nucleatum GroEL with HMEC-1 cells and subsequently induces an inflammatory response via the MAPK and NF-κB pathways.

소음 스트레스로 인한 뱀장어(Anguilla japonica)의 영향을 파악하기 위하여 스트레스 지표로 사용되는 코티졸, 포도당, 알부민 및 glucocorticoid receptor(GCR) 유전자의 발현 양을 측정, 분석하여 노출되지 않은 대조구와 비교하였다. 그 결과, 알부민은 노출 1시간 후에 낮은 값을 보인 반면 코티졸과 포도당은 대조구에 비해 매우 큰 차이를 보이며 높게 나타났다. GCR 유전자의 조직 발현 결과 간, 아가미, 근육 및 소장에서 많이 발현하였다. 소음 노출에 따른 시간의 변화에서 간과 아가미 근육과 소장에서 발현이 감소하는 양상을 나타내었다. 실험결과 뱀장어의 glucocorticoid receptor 유전자의 발현변화가 소음 스트레스로 인한 영향을 파악하는데 유용한 지표가 될 수 있음을 확인하였다.

Urokinase-type plasminogen activator(uPA) bound to urokinase plasminogen activator receptor(uPAR) expression is strongly correlated with the metastatic potential of various tumors by enhancing ECM degradation through plasminogen and matrix metallopreotease activation. But expression of uPA/uPAR in human malignant salivary gland tumors has been rarely reported. The purpose of this study were to investigate mRNA expression and cytologic concentration of uPAR in SGT cell line compared to various cancer cell lines by RT-PCR and ELISA method, and to study migration and adhesion assay. These results would be to apply the pathogenesis and prognosis of malignant salivary gland tumors. All the cell lines(SGT, HN 4, SCC 25, and HeLA) were cultured under DMEM with 10% FBS at 37℃ in a 5% CO2 incubator. We studied a possible association between mRNA expression and cytosolic concentrations of uPAR in SGT cell line compared to various cancer cell lines using RT-PCR and an enzyme-linked immunoassay( ELISA) method. And also cell adhesion and migration assay were done in all the cell lines. In migration assay SGT cell line was about 2.5-4 folds higher than another cell lines. In adhesion assay SGT cell line was about 1.1-2 folds higher than another cell lines. uPAR cytosolic concentrations of SGT cell line was about 3.4-10 folds by ELISA, while mRNA expression was about 2.5-5 folds by RT-PCR. Oral Scc cell lines showed the lowest value. uPAR protein and mRNA expression were correlated with migration and adhesion assay. From the aboving results, high cytosolic concentrations and mRNA expression of uPAR were correlated with migration and adhesion assay. It suggested that this might be a specific marker for malignant potential of SGT cell line and would be contributed to pathogenesis and prognosis of human salivary gland adenocarcinoma

β-phenylethyl isothiocyanate(PEITC) is a component derived from cruciferous vegetables and has been demonstrated to fight many types of cancers through various molecular pathways. In the present study, we focused on its effect on the induction of apoptotic cell death to inhibit cell growth and its molecular mechanism in HSC-4 human oral cancer cells. A colorimetric MTS assay was used to examine cell viability. The apoptotic effect and was investigated using DAPI staining and the molecular target and mechanism of PEITC-mediated apoptosis were determined by Western blotting. The result showed that PEITC inhibited oral cancer cell growth and induced apoptosis via extrinsic signaling pathway evidenced by the activation of caspase 8, truncation of bid protein and induction of death receptor(DR) 5. DR5 protein level was increased through the activation of p38 and c-Jun N-terminal kinase(JNK). These results from this study strongly suggest that DR5 is a potential molecular target for PEITC-induced apoptosis in oral cancer via p38 and JNK.

The purpose of this study was to investigate the body composition, biochemical parameters, and consumption of convenience foods according to β-3 adrenergic receptor polymorphism in university students. A survey was conducted on a total of 486 students - 189 males and 297 females. Based on a self-reporting method, questionnaires were administered for over 20 minutes, and β-3 adrenergic receptor and blood samples were also analyzed. The genotype frequencies of β-3 adrenergic receptor polymorphism were Trp/Trp homozygote (73.0%) and Trp/Arg heterozygote (27.0%) in male students. For the female students, the distribution of genotypes was Trp/Trp (71.0%) and Trp/Arg (29.0%). There were no differences according to biochemical parameters (ALT, cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol, and hemoglobin) or body composition. Males with TT genotype frequently ate Ramyon (2.40±0.52), Cup Ramyon (2.37±0.39), Kimchi (2.23±0.61), and frozen meat (2.00±0.44), whereas males with TA genotype ate Fries (frozen food) (1.90±0.79), Smoked meat (1.67±0.81), and Canned fruit (1.64±0.81). Females with TT genotype frequently ate Frozen fries (2.21±0.35), Kimbab (2.12±0.44), and Ramyon (1.85±0.40), whereas females with TA genotype frequently ate Kimchi (1.73±0.98), Fries (frozen food) (1.46±0.26), and Cup Ramyon (1.30±0.34). When questioned about satisfaction about body shape, 22.8 and 60.8% of those with TT genotype answered that they were 'satisfied' or needed to 'lose weight', respectively, whereas 18.0 and 63.9% of those with TA genotype answered that they were 'satisfied' or needed to 'lose weight', respectively. In conclusion, this study found no significant effects in terms of β-3 adrenergic receptor polymorphism, which suggests that health-promoting education needs to be developed so that university students appropriately recognize their bodies and control their weight in desirable ways. Therefore, it is necessary to educate individuals with TT genotype how to buy reasonable foods by understanding the interrelationship between convenience foods and health care and by checking the nutrition index labels on convenience foods. Thus, it is recommended that a health-promoting program be developed for the promotion of healthy lifestyles.

Scavenger receptors (SRs) are transmembrane cell surface molecules recognized in apophotic cells, bacteria and lipopolysaccharide. With no physiological information on SRs in insects except SR-CI of Drosophila melanogaster, a putative SR gene was cloned and characterized in Spodoptera exigua. A partial S. exigua SR gene was obtained from hemocyte transcripts and exhibited high homology with type C. Its expression was confirmed in all developmental stages. Among different tissues, S. exigua SR was expressed highly in hemocytes. To confirm change in SR expression by infection, Escherichia coli was injected to fifth instar and RNA was extracted after 10 hours. SR expression in hemocytes of E. coli injected larva was not significantly different from the control but SR expression in fat body of E. coli injected larva was higher than the control. It is expected that SRs of S. exigua are related with immune responses against bacteria such as E. coli. To address its function, S. exigua SR expression was suppressed by double-stranded RNA (dsRNA).

To search the potent pig pheromonal odorants through receptor-based approach methods, molecular dockings between 680 Flavornets as substrate molecule and pig odorants binding proteins OBP (1HQP) and PBP (1GM6) as receptor, and QSPR (quantitative structure-property relationship) analyses from physico-chemical parameters of Flavornets and their docking scores (DS) were performed and discussed quantitatively. From the basis on the findings, the optimal value (MSA)opt.=407.595 Å2 of MSA (molecular surface area; Å), and RB (number of rotational bond) had the Flavornets will be able to increase DS. Therefore, it is expected that the stearyl alcohol from DS and H-bond type between substrate and receptor would be shows the character as potent pig pheromonal odorant.

Upon mating, females of many animal species undergo dramatic changes in their behavior. In Drosophila melanogaster, post-mating behaviors are triggered by sex peptide (SP), a key modulatory substance produced in the male seminal fluid and transferred to female during copulation. SP modulates female behaviors by acting on the sex peptide receptor (SPR) located in a small subset of internal sensory neurons that innervate the female uterus and project to the central nervous system (CNS). Interestingly, however, SPR is also expressed broadly in the CNS of both sexes. Moreover, SPR is also encoded in the genomes of insects that lack obvious SP orthologs. Based on these observations, we speculated that SPR may have additional ligands that are only distantly related to SP, if at all. If so, then this also raises questions on the evolution of SP-SPR signaling. To begin to address these questions, we set out to identify additional ligands for SPR. Here, we identify myoinhibitory peptides (MIPs) as a second family of SPR ligands that is conserved across a wide range of invertebrate species. MIPs are potent agonists for Drosophila, Aedes and Aplysia SPRs in vitro, yet are unable to trigger post-mating responses in vivo. In contrast to SP, MIPs are not produced in male reproductive organs, and are not required for post-mating behaviors in Drosophila females. We conclude that MIPs are evolutionarily conserved ligands for SPR, which are likely to mediate functions other than the regulation of female reproductive behaviors. Therefore, we propose that SPR has a different ancestral function, with a role in post-mating behavior arising only recently in Drosophila evolution, concomitant with the emergence of its novel SP ligand.

Medial vestibular nucleus (MVN) neurons are involved in the reflex control of the head and eyes, and in the recovery of vestibular function after the formation of peripheral vestibular lesions. In our present study, whole cell patch clamp recordings were carried out on MVN neurons in brainstem slices from neonatal rats to investigate the actions of a group I metabotropic glutamate receptor (mGluR) agonist upon synaptic transmission and ionic currents. Application of the mGluR I agonist (S)-3,5- dihydroxyphenylglycine (DHPG) increased the frequency of miniature inhibitory postsynaptic currents (mIPSCs) but had no effect upon amplitude distributions. To then identify which of mGluR subtypes is responsible for the actions of DHPG in the MVN, we employed two novel subtype selective antagonists. (S)-(+)--amino-a-methylbenzeneacetic acid (LY367385) is a potent competitive antagonist that is selective for mGluR1, whereas 2-methyl-6-(phenylethynyl)-pyridine (MPEP) is a potent noncompetitive antagonist of mGluR5. Both LY367385 and MPEP antagonized the DHPG-induced increase of mIPSCs, with the former being more potent. DHPG was also found to induce an inward current, which can be enhanced under depolarized conditions. This DHPG-induced current was reduced by both LY367385 and MPEP. The DHPG-induced inward current was also suppressed by the PLC blocker U-73122, the IP₃ receptor antagonist 2-APB, and following the depletion of the intracellular Cα2+ pool by thapsigargin. These data suggest that the DHPG-induced inward current may be mainly regulated by the intracellular Cα2+ store via the PLC-IP3 pathway. In conclusion, mGluR I, via pre- and postsynaptic actions, may modulate the excitability of the MVN neurons.

A cDNA of PBAN receptor (Plx-PBANR) isolated from female pheromone gland of the diamondback moth encodes 338 amino acids and has 7 transmembranes, belonging to G-protein coupled receptor family. The fact that Plx-PBANR expression was only found in female pheromone gland revealed that pheromone gland is the only molecular target of Plx-PBAN. Plx-PBANR expressing cells increased level of Ca2+ influx when challenged with PBANs. When RNAi fragment for PBANR was injected into pupae, suppression of PBANR expression was maintained for at least 2 days at post-emergence. Injection of RNA fragment for inhibition of Plx-PBANR expression also inhibited mating behavior and suppressed sex pheromone production, suggesting that some molecular target was affected by reduced Plx-PBANR expression. We cloned partial Δ9 and Δ11 desaturase gene and investigated expression level in Plx-PBANR-RNAi moth. It is of interest that desaturases expression was reduced by RNA fragment injection. These results suggest of PBANR expression affects the molecular biological events of PBAN and eventually suppresses mating behavior.