Bee venom contains a variety of peptide constituents that have various biological, toxicological, and pharmacological actions. However, the biological actions of secapin, a venom peptide in bee venom, remain largely unknown. Here, we provide the first evidence that the Asiatic honeybee (Apis cerana) secapin (AcSecapin-1) exhibits anti-fibrinolytic, anti-elastolytic, and anti-microbial activities. AcSecapin-1 functions as a serine protease inhibitor-like peptide that has inhibitory effects against plasmin, elastases, microbial serine proteases, trypsin, and chymotrypsin. Consistent with these functions, AcSecapin-1 inhibited the plasmin-mediated degradation of fibrin to fibrin degradation products, thus indicating the role of AcSecapin-1 as a clotting factor. AcSecapin-1 also inhibited both human neutrophil and porcine pancreatic elastases. Furthermore, AcSecapin-1 exhibited anti-microbial activity against fungi and Gram-positive and Gram-negative bacteria. Taken together, our data demonstrated that AcSecapin-1 has a multifunctional role as an anti-fibrinolytic agent, an anti-elastolytic agent, and an anti-microbial peptide, and our data suggested novel functions for the biological actions of the bee venom peptide, secapin.

The lycanid butterfly, Shijimiaeoides divina (Lepidoptera: Lycaenidae) is listed as the second-degree endangered wild animal in Korea from 2012. The 15,259-bp long complete mitochondrial genome of the species consisted of a typical set of genes (13 protein-coding genes, two rRNA genes and 22 tRNA genes) and one major non-coding A+T-rich region, with the typical arrangement found in majority of Lepidoptera. The 15,259-bp long S. divina mitogenome is well within the range found in Lycaenidae and has typical sets of 37 genes and a major non-coding A+T-rich region as 379 bp. As other lycanid butterflies S. divina COI also started with CGA. The gene arrangement of S. divina is identical to that of the Ditrysia in Lepidoptera that has the order trnM-trnI-trnQ (underline for inverted gene) between the A+T-rich region and ND2. Comparison of the skewness between the PCGs encoded in major and minor strand indicates a substantial difference between them in GC skewness (0.261 ~ 0.340 in minor strand vs. -0.081 ~ -0.115 in major strand). The 151-bp intergenic spacer sequence of the S. divina mitogenome is spread over 16 regions ranging in size from 1-53 bp. The longest one (53 bp) located between trnQ and ND2 shows substantially high sequence homology to neighboring ND2 may indicating the origination of the region by a partial duplication of the ND2 gene. One of the unusual features of the S. divina mitogenome is the presence of a trnK-like sequence that is encoded at the major strand of the genome in the A+T-rich region.

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.

This studies was conducted to define community structure and diversity of epigeal spiders in apple orchards under different farming practices (organic and conventional). In 11 commercial apple orchards, epigeal spider community structure was investigated during 2012-2014 (3 orchards, 2012; 3 orchards, 2013, 5 orchards, 2014). Epigeal spiders were collected by pitfall traps from April to October, 2012 in Danyang-gun, Chungcheongbuk-do; 2013 in Muju-gun, Jeollabuk-do; 2014 in Cheongsong-gun and Uiseong-gun, Gyeongsangbuk-do, Korea. Sampled spiders were identified to the species level. Biodiversity was analyzed with species richness (total numbers of spider species), abundance (total numbers of individuals) and Shannon-Wiener diversity index (H΄) for each site and compared between organic and conventional orchards. In all regions, abundance of the spider communities were relatively higher in organic orchards than in conventional orchards. But, no significant difference in species richness and specie diversity (H΄) between organic and conventional orchards. Lycosidae was the most dominant family in all orchards regardless farming practices. However, Linyphiidae dominated in organic orchards only. The most dominant species were Pardosa laura (Lycosidae), Erigone koshiensis (Linyphiidae) in organic, Piratula procurvus (Lycosidae) in conventional orchards.

Insect and natural enemy fauna were surveyed at conventional (CV) and organic-farming persimmon orchards (OF) at Jinju, Korea, using sweeping net and naked eyes in 2013 and 2014. Using sweeping net, 49 species at an OF and 18 species at a CV were observed in 2013. In 2014 too, more species were observed at OF (66 species, 33 families, 7 orders) than at a CV (25 species, 15 families, 5 orders). During both the years, dominant species were all hemipteran insects like Nysius plebejus (Lygaeidae) and Cletus punctiger (Coreidae) in both types of orchards, followed by Rhopalus maculatus (Rhopalidae), Riptortus pedestris (Alydidae). Similarly, naked eye inspection also found more species at OF (192 species, 72 families, 11 orders) than at CV (104 species, 52 families, 10 orders) in 2014. Dominant species in naked eye inspection were Lycorma emelianovi (Hemiptera: Fulgoridae), Apis mellifera (Hymenoptera: Apidae), Uroleucon cephalonopli (Hemiptera: Aphididae) at OF, and A. mellifera and R. maculatus at CV in 2014. Using naked eye inspection or sweeping net in 2014, more species of natural enemies were observed at OF (31 and 9 species, respectively) than at CV (18 and 5 species, respectively).

Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide.

We have used bulked segregant analysis to screen the strain-specific DNA marker associated thermophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 550 bp was amplified with OP-S07 primer from the thermophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-S07-1. The PCR analysis with the OP-S07-1 primer showed that this SCAR marker clearly distinguish the thermophilic strains from the control strains.

This study was conducted to investigate the community structure and species distribution of ground beetle assemblages in western part of Yeongwol-gun, Korea. Ground beetles were collected by pitfall traps from 9 study sites during July to October in 2013. A total of 30 species were identified from 4,439 collected ground beetles. Two dominant species, Synuchus nitidus (1,916 individuals) and Synuchus cycloderus (1,900 individuals) were occupied 85.8% of total. Interestingly, Pterostichus orientalis orientalis and Synuchus melantho were only collected in relative high altitude above 350 m, such as Beopheung-ri, Unhak-ri, and Yongseok-ri. Large-sized beetles, such as Damaster, Leptocarabus, and Carabus species, were generally more collected from Suju-myeon (Beopheung-ri and Unhak-ri) and Jucheon-myeon (Docheon-ri and Yongseok-ri) as well-conserved area, but not occurred in Yeongwol-eup and Hanbando-myeon, which forests may have lower habitat quality for these species. These differences also lead to increasing of dissimilarity among study sites, which were diverged into 3 groups at 53.0% of similarity. In particular, species composition of Samok-ri was significantly different at 8.2% of similarity because of difference in habitat characteristics as river side. This study has an important value to provide the fundamental information of inventory and monitoring in Yeongwol-gun. For detail mapping of species distribution in Yeongwol-gun, however, further studies are necessary, especially eastern parts of Yeongwol-gun.

Epigeic spiders (Araneae) were surveyed in apple orchards under different farming practices (organic and conventional) in Muju-gun, Jeollabuk-do, Korea. The study was conducted from April to October in 2013 with 2 weeks interval using pitfall traps. Collected spiders were identified as 20 species of 6 families from 819 individuals in organic orchardⅠ, 19 species of 10 families from 759 individuals in orchardⅡ and 16 species of 8 families from 209 individuals in conventional orchard for from 1,787 captured spiders. Biodiversity was analyzed with species richness, abundance and Shannon diversity index (H΄). Species richness was statistically higher in organic orchard than in conventional orchard. Organic orchardⅡ comprised the greatest abundance followed by organic orchard and conventional. But species diversity was the highest in organic orchardⅡ, intermediate in conventional orchard and the lowest in orchardⅠ. Lycosidae was the most dominant family in all orchards regardless farming practices. However, Linyphiidae dominated in organic orchards only. The most dominant species were Erigone koshiensis (Linyphiidae) in organicⅠ, Pardosa laura (Lycosidae) in organicⅡ and Arctosa pungcheunensis (Lycosidae) in conventional orchard.

Aggregatibacter actinomycetemcomitans is an important pathogen in the development of localized aggressive periodontitis. Lipopolysaccharide (LPS) is a virulent factor of periodontal pathogens that contributes to alveolar bone loss and connective tissue degradation in periodontal disease. Our present study was designed to investigate the cytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine gene expression profiling in RAW 264.7 cells was performed by microarray analyses. The cytokine mRNA and protein levels and related signaling pathways induced by Aa LPS were measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly induced the expression of NF-κB, NF-κB-related genes, inflammatory cytokines, TNF-α and IL-1β in RAW 264.7 cells. NF-κB inhibitor pretreatment significantly reduced the levels of TNF-α and IL-1β mRNA and protein. In addition, the Aa LPS-induced TNF-α and IL-1β expression was inhibited by p38/JNK MAP kinase inhibitor pretreatment. These results show that Aa LPS stimulates TNF-α and IL-1β expression through NF-κB and p38/JNK activation in RAW 264.7 cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.

Insect cuticular melanization is regulated by the prophenoloxidase (proPO)- activating system, which is also involved in the innate immune reaction. Here, we demonstrate how the differentiation of the proPO-activating system is regulated toward a cuticular melanization or innate immunity function in silkworm (Bombyx mori) pupae. Our results indicate that the differential and spatial regulation of key components, such as the proPO-activating factor, tyrosine hydroxylase, and porPOs, primes the proPO-activating system for either cuticular melanization or innate immunity. This dual strategy for cuticular melanization in development and innate immunity upon infection demonstrates a two-pronged defense mechanism that is mediated by the priming of the proPO system.

Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or typsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, B. thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.