A new fumigant, carbonyl sulfide (COS), has potential for use as a replacement for methyl bromide, yet its mechanism of toxicity to insects remains poorly understood. In this study, transcriptome analysis was performed on Tribolium castaneum malpighian tubules and fat bodies, which are known to play an essential role in energy storage and utilization in insect species. In total, upon exposure to COS, 3,034 and 2,973 genes were differentially expressed in the T. castaneum malpighian tubules and fat body, respectively. These differentially expressed genes comprise a significant number of detoxification-related genes, including 105 P450s, 18 glutathione S-transferases (GSTs), 82 ABC transporters, 25 UDP-glucosyltransferases and 42 carboxylesterases and mitochondrial–related genes, including 9 complex Ⅰ genes, 2 complex Ⅱ genes, 1 complex Ⅲ gene, 9 complex IV genes, 8 complex V genes from both malpighian tubules and fat body tissues. Moreover, KEGG analysis demonstrated that the upregulated genes were enriched in xenobiotic metabolism by ABC transporters and drug metabolism by other enzymes. We also investigated the role of carbonic anhydrases (CAs) in toxicity of COS using dsRNA treatment in T. castaneum. These results show that CA genes have a key role in toxicity of the COS. Furthermore, the results of transcriptomic analysis provide new insights into the insecticidal mechanism of COS fumigation against T. castaneum and eventually contribute to the management of this important stored grain pests.

Short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate are secondary metabolites produced by anaerobic fermentation of dietary fibers in the intestine. Intestinal SCFAs exert various beneficial effects on intestinal homeostasis, including energy metabolism, autophagy, cell proliferation, immune reaction, and inflammation, whereas contradictory roles of SCFAs in the oral cavity have been reported. Herein, we found that low and high concentrations of SCFAs induce differential regulation of intracellular Ca2+ mobilization and expression of pro-inflammatory cytokines, such as interleukin (IL)-6 and IL-8, respectively, in gingival fibroblast cells. Additionally, cell viability was found to be differentially regulated in response to low and high concentrations of SCFAs. These findings demonstrate that the physiological functions of SCFAs in various cellular responses are more likely dependent on their local concentration.

Exosomes are Nano-sized lipid vesicles secreted from mammalian cells containing diverse cellular materials such as proteins, lipids, and nucleotides. Multiple lines of evidence indicate that in saliva, exosomes and their contents such as microRNAs (miRNAs) mediate numerous cellular responses upon delivery to recipient cells. The objective of this study was to characterize the different expression profile of exosomal miRNAs in saliva samples, periodically isolated from a single periodontitis patient. Unstimulated saliva was collected from a single patient over time periods for managing periodontitis. MicroRNAs extracted from each phase were investigated for the expression of exosomal miRNAs. Salivary exosomal miRNAs were analyzed using Affymetrix miRNA arrays and prediction of target genes and pathways for its different expression performed using DIANA-mirPath, a web-based, computational tool. Following the delivery of miRNA mimics (hsa-miR-4487, -4532, and -7108-5p) into human gingival fibroblasts, the expression of pro-inflammatory cytokines and activation of the MAPK pathway were evaluated through RT-PCR and western blotting. In each phase, 13 and 43 miRNAs were found to be differently expressed (|FC| ≥ 2). Among these, hsa-miR-4487 (|FC|=9.292005) and hasmiR- 4532 (|FC|=18.322697) were highly up-regulated in the clinically severe phase, whereas hsa-miR-7108-5p (|FC|= 12.20601) was strongly up-regulated in the clinically mild phase. In addition, the overexpression of miRNA mimics in human gingival fibroblasts resulted in a significant induction of IL-6 mRNA expression and p38 phosphorylation. The findings of this study established alterations in salivary exosomal miRNAs which are dependent on the severity of periodontitis and may act as potential candidates for the treatment of oral inflammatory diseases.

β-glucan is a safe and highly potent biological response modifier that nutritionally activates the immune response through the Macrophage, Dendritic and additional immune cells to yield various therapeutic effects. Shiitake mushrooms (Lentinula edodes) contanining β-glucans may be beneficial for human health; they have been used in the treatment of cancer, hypertension, and high cholesterol levels. The effect of different substrates and various developmental stages (mycelium growth, primordium appearance, and fruiting-body formation) on β-glucan production in the edible mushroom L. edodes was studied. The cap of the mature mushroom showed the highest β-glucan activity, and β-glucan activity seemed to be influenced somewhat by some well-known inducers or sawdust. In this study, we utilized five strains (JMI 10022, 10036, 10077, 10079, 10080) of L. edodes regardless of origin and growth conditions. This experiment showed that the expression of β-glucan was induced by glucose bond, and increased with the growing of L. edodes. Quantitatively, reverse transcription PCR utilized pairs of primers specific for β-glucan gene expression shows that expressed genes were most commonly indentified during the process of fruiting-body formation. We suggest that the results will provide valuable information to assist L. edodes industry.

Salt stress is one of the most limiting factors that reduce plant growth, development and yield. However, identification of salt-inducible genes is an initial step for understanding the adaptive response of plants to salt stress. In this study, we used an annealing control primer (ACP) based GeneFishing technique to identify differentially expressed genes (DEGs) in Italian ryegrass seedlings under salt stress. Ten-day-old seedlings were exposed to 100 mM NaCl for 6 h. Using 60 ACPs, a total 8 up-regulated genes were identified and sequenced. We identified several promising genes encoding alpha-glactosidase b, light harvesting chlorophyll a/b binding protein, metallothionein-like protein 3B-like, translation factor SUI, translation initiation factor eIF1, glyceraldehyde-3-phosphate dehydrogenase 2 and elongation factor 1-alpha. These genes were mostly involved in plant development, signaling, ROS detoxification and salt acclimation. However, this study provides new molecular information of several genes to understand the salt stress response. These genes would be useful for the enhancement of salt stress tolerance in plants.

The coat color of mammals is determined by the melanogenesis pathway, which is responsible for maintaining the balance between black-brown eumelanin and yellow-reddish pheomelanin. The melanogenesis-associated genes controlling pigmentation act as a complex and interact with each other to cause phenotypic and genotypic variations in cattle. That the MC1R genotype of Korean native cattle with dark muzzle was e/e or E+/e, while the genotype of Korean native cattle with light muzzle was E+/E+, which is a variant of the MC1R genotype in the Korean native cattle. Especially, the MC1R expression type is shows how much pigmentation, important factor in deciding its status in the coat and nose colours. However, information regarding the coat or nose colours-associated gene regulation of korean cattle is not yet unknown. Therefore, in this study was to investigate the expression patterns of melanogenesis-associated genes in black dot nose(korea brindle cattle) and normal nose(korea native cattle). Using microarray clustering and real-time polymerase chain reaction techniques, we analysed that the expression of genes involved in the mitogen-activated protein kinase (MAPK) and Wnt signaling pathways is distinctively regulated in the dark and light muzzle tissues. Differential expression of tyrosinase was also noticed, although the difference was not as distinct as those of MAPK and Wnt. We hypothesize that emphasis on the MAPK pathway in the Korea brindle cattle induces eumelanin synthesis through the activation of cAMP response elementbinding protein and tyrosinase, while activation of Wnt signaling counteracts this process and raises the amount of pheomelanin in the native cattle. Regarding the increasing interest in the genetic diversity of cattle stocks, genes we identified for differential expression in the brindle cattle vs. native cattle may serve as novel markers for genetic diversity among cows based on the coat and muzzle color phenotype.

The small brown planthopper (SBPH), Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus (RSV) which often causes significant reduction of yield in the field. Buprofezine is an effective insect growth regular (IGR) pesticide to control planthoppers, however, since the use of buprofezine for more than a decade, it has caused a certain resistance of SBPH. To survey the responses of SBPH to buprofezine, we exposed 4th instar SBPH to 200 ppm buprofezine by dipping method, and extracted total RNA for RNA-seq by Illumina platform. The quality filtered raw reads of cDNA obtained from experimental and control SBPH were subjected to Bowtie2 followed by eXpress computer program to compare the differential gene expression which will be important information for pest control methods using RNAi.

Tooth development shows dynamic morphological changes from the stages of cap to hard tissue formation and is strictly regulated during development. In the present study, we compared expression and localization of 3 major enamel matrix proteins in rats: amelogenin, enamel and ameloblastin. DD-PCR and RT-PCR revealed differential expression of the major proteins from the cap stage to root stage. Immunofluorescence staining results indicated that amelogenin was not detected in either inner enamel epithelium or reduced enamel epithelium, but highly immunoreactive in preameloblasts and ameloblasts; in addition, it was sporadically expressed in preodontoblasts abutting preameloblasts. Ameloblastin expression was also observed in not only differentiated ameloblasts but also osteoblasts. Immunoreactivity to ameloblastin in ameloblasts was strong in Tomes' processes. Enamelin was exclusively localized along the entire newly formed and maturing enamel. Enamelin was largely localized in near Tomes' processes and enamel rods in maturing enamel. Alendronate treatment resulted in down-regulation of amelogenin and ameloblastin at both transcription and translation levels; whereas, enamelin expression was unchanged in response to the treatment. These results suggested that amelogenin, ameloblastin and enamelin might be implicated in cell differentiation, adhesion of ameloblasts to enamel and enamel crystallization during enamel matrix formation, respectively.

The tongue has 4 kinds of papillae, which are filiform, fungiform (FU), foliate (FO) and circumvallate papilla (CV). Tongue papillae except filiform papilla include taste buds. The papillae differ in taste sensitivities, likely due to differential expression of taste receptors. In this study, we evaluated differences in the expression levels of taste receptors in FU, FO and CV. Male DBA2 mice, 42-60 days old, were used in the study. Messenger RNAs were extracted from the murine epithelial tissues including FU, FO and CV. Cloned DNAs were synthesized by reverse transcription. Quantitative PCRs (qPCRs) were performed to determine mRNA expression levels of taste receptors. Results of qPCR revealed that the relative expression levels and patterns were different among FU, FO and CV. All three type 1 taste receptors were expressed FU, FO and CV at varying relative expression levels. All 35 kinds of type 2 taste receptors showed higher expression in FO and CV than in FU. Tas2r108 and Tas2r137 showed the two highest expression levels in all tested papillae. The differential expression levels and patterns of taste receptors among the three papillae could contribute to the different physiological sensitivities by tongue areas. Additional studies such as in situ hybridization or taste receptor cell activity recording is necessary to elucidate the functional relationship between expression levels of taste receptors and taste sensitivity.

Insect Growth Regulators (IGRs) are insecticides that disrupt the normal development of target insects by inducing symptoms such as premature molting or supernumerary larval stages. Juvenile hormone systems become the targets of two types of IGRs: the Juvenile Hormone Agonists (JHAs) and Juvenile Hormone Antagonists (JHANs). Pyriproxyfen is one of the chemical compounds widely used as JHA to control many kinds of insects while Kanakugiol is a plant-extracted compound which acts as JHAN. The small brown planthopper, Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus which often causes significant reduction of yield in the field. In order to analyze the differential gene expressions of L. striatellus upon JHA and JHAN treatment by using next generation sequencing technique, we sprayed Pyriproxyfen and Kanakugiol on 4th instar nymphs of L. striatellus respectively, and extracted total RNA for RNA-seq. The quality-filtered Illumina sequence reads of the control, JHA, and JHAN treated samples were mapped to the reference gene sequences by using the Bowtie2 software. Then the results of mapping by Bowtie2 were analyzed by eXpress software to quantity the differential gene expression.

Five heat shock protein 70 (hsp70) isoforms (hsc70-3, hsc70-4, hsc70-5, hsc70cb, hsp70Ab) of Apis mellifera were identified from Honeybee genome database. Specific primers for each isoform were designed for the quantitative realtime RT-PCR analysis, then analyzed transcript levels of the abdomen of adult workers (3-4 weeks old) in respond to heat shock and imidacloprid ingestion. Heat shock at 45°C for 1 h induced all 5 genes but highest in hsc70cb, hsp70Ab. Ingestion of imidacloprid pesticide did not change any hsp70 isoforms at 33°C but those bees also highly responsive to heat shock at 45°C. In addition, expression level of each hsp70 isoform was various and heat shock response of each isoform was tissue-specific. For example, hsc70-3 was highest in midgut, hsc70-4 was in hypopharyngeal gland, but hsp70Ab was in fat bodies. However, heat shock response of all 5 isoforms was the highest in the fat body than brain, hypopharyngeal gland, midgut, flight muscle and integument. Our results provide information for the understanding of complicated protective mechanism of honey bee against thermal stress.

Lysyl oxidase (LOX) family, the copper dependent amine oxidase, oxidizes lysine residues in extracellular collagen and elastin. LOX increases the strength of the extracellular matrix and plays an important role in tumor development and metastasis. It has been reported that increased LOX protein and RNA are found in head and neck squamous cell carcinoma. Moreover some studies regarded LOX as a prognostic marker of oral and oropharyngeal squamous cell carcinoma. However there has not been any report on LOX expression of salivary gland tumors. Here, we investigated LOX expression in mucoepidermoid carcinoma (MEC) and adenoid cystic carcinoma (ACC) of salivary gland and compare it to those of pleomorphic adenoma (PA). We evaluated LOX expression in eighteen MEC, eighteen ACC and twenty PA cases by immunohistochemical examination. Whereas PA showed relatively low density of LOX expression, ACC revealed more cases that showing high staining intensities for LOX. Significantly increased LOX expression was found in the cases of ACC when compared to those of PA (P = 0.010).

This experiment was conducted to investigate the genetic effects of candidate genes on the growth of spleen and liver tissues using dietary Curcuma longa (C. longa) supplementation. Expression analyses of candidate genes regarding animal growth was performed in order to determine the factors affecting the growth related to immune components of Curucumin, Turmerone, and Zingiberene as the bile secretion Paratolyl methyl carbinol (PTMC). The animals were divided into four groups of five chicks supplied with experimental diets of C. longa at 0.25, 0.5 and 1% and controls. The 19 growth-related genes were known to cell maturation, differentiation significant expression patterns in this analysis. Expression of growth response-related genes in chicks supplemented with 1% of C. longa showed better growth performance than chicks with 0.25 and 0.5% in spleen (p<0.05). The IGF1, MSTN, POU1F1, ADCYAP1 gene were known to central roles in mediating gonadotropin function, regulating steroidogenesis and promoting oocyte growth and maturation. Sex steroids, androgen and estrogen can affect sex differentiation and also can affect muscle development. On the other hand, GHSR and FABP3 gene showed significant expression patterns in this analysis. The results would be used as basic information for the variation of growth-related genes expression on the cell growth, sex cell growth, and sex hormones according to dietary supplementation with C. longa in chickens.

국내야생종인 새머루로부터 내병성 포도품종 육성에 필요한 육종소재와 유용정보를 획득하고자, 포도 새눈무늬병균과 줄기혹병균에 감염된 새머루에서 식물 방어관련유전자의 발현양상을 분석하였다. Thaumatin (TLP), glutathione peroxidase (GPX), glutathione-S-transferase (GST), chalcone synthesis (CHS), protein kinase regulator (14-3-3) 및 β-glucanase (Glu) 등의 방어관련유전자, leucine reach-repeat (LRR)와 lipoxygenase (LOX) 등의 신호전달관련유전자, polygalacturonase-inhibiting protein (PGIP)과 같은 세포벽관련 유전자의 발현이 포도 새눈무늬병균 접종에 의해 유도되었다. 포도새눈무늬병균의 포자접종 및 배양여액처리에 의해 WRKYtranscription factor 10 (WRKY), fatty acid elongase (FAE)의 발현이 유도되었으며 proline-rich protein (PRP) 유전자의발현은 억제되었다. 발현이 증가한 15개 유전자 중에서 LRR, LOX, TLP, GST 등은 포도새눈무늬병균 포자현탁액 접종에 의해 발현이 크게 증가하였으나, CHS와 tonoplast intrinsicprotein (TIP)는 배양여액처리에 의해 증가하였다. 포도 줄기혹병균의 접종에 의해 LRR, CLP, LOX, TLP, GPX, 14-3-3,GST, PGIP, FAE, TIP, Glu, WRKY 등은 발현이 증가하였고, CHS와 PRPs는 발현이 억제되었다. 포도 줄기혹병균 접종에 의해 활성산소의 축적 및 분해와 관련된 GPX와 GST는 발현이 크게 증가하였다.

Tooth development involves bud, cap, bell and hard tissue formation stages, each of which is tightly controlled by regulatory molecules. The aim of this study was to identify genes that are differentially expressed during dental hard tissue differentiation. Sprague-Dawley rats at postnatal days 3, 6 and 9 were used in the analysis. Differential display RT-PCR (DD-PCR) was used to screen differentially expressed genes between the 2nd (root formation stage, during mineralization) and 3rd (cap stage, before minerali-zation) molar germs at postnatal day 9. The DNA detected in the 2nd molar germs showed homology to osteonectin only (GenBank accession no. NM_012656.1). The level of osteonectin mRNA expression was much higher in the 2nd molar germs than in the 3rd molar germs and was found to increase in a time-dependent manner from the early bell stage to the root formation stage in the 2nd molar germs. The pattern of osteonectin protein expression was consistent with these RT-PCR results. Osteonectin protein was found by immunofluorescent analysis to localize in odontoblasts and preodontoblasts rather than the dentin matrix itself. Further studies are needed to validate the involvement of osteonectin in mineralization and root formation.

Pleomorphic adenoma (PA) is the most common benign salivary gland tumor. It is biphasic and is characterized by an admixture of epithelial and spindle-shaped myoepithelial cells in a variable background stroma. Epithelial-myoepithelial carcinoma (EMC) is a malignant biphasic salivary gland tumor typically composed of clear myoepithelial cells that surround epithelial-lined ducts resembling intercalated ducts. The differential diagnosis between the two tumor may be occasionally encountered because of the shared histophatologic feature. And then, it would be more reliable to differentiate the tumors based on biological behavior such as the expression of distinct intermediate filaments such as cytokeratin, invasiveness- related molecules, and the growth factor receptor to aberrantly facilitate the tumor growth, and the growth fraction of tumors. Therefore, from the 10 cases of PA and 6 cases of EMC, we immunohistochemically examined the differential expression of the cytokine 7 and 14, matrix metalloproteinase-9, C-KIT, and Ki-67 between the two tumor. At the results, there were significant differences of CK7 expression in non-luminal cells (P = 0.000) and CK14 expression in luminal and non-luminal cells of the both tumors (P = 0.025 and P = 0.000, respectively). In the comparison of the biologic behavior, a significantly increased expression of MMP-9, C-KIT and Ki-67 was found in the cases of EMC when compared to those of PA (P = 0.043, P = 0.011, and P = 0.000, respectively). In conclusion, the differences of CK expression in luminal and non-luminal cells between PA and EMC seem to reflect the difference of the origin and the level of the maturation of the tumor cell. Increased expression of MMP-9, C-KIT, and Ki-67 in EMC may represent more aggressive biologic behavior of the tumor compared with benign salivary tumor such as PA. Our results may be helpful to understand the histiogenesis of the two tumors and the difference of biologic behavior and to differentiate them when the limited specimen was submitted. Further study of many more cases of EMC is needed to validate the usefulness of these molecules as the diagnostic aid.