To identify and compare the venom components and expression patterns of some bees/wasps, venom gland-specific transcriptome analyses were conducted for 14 Aculeate bees/wasps. Most of the allergens and pain-producing factors showed extremely high expression levels in social wasps, implying that social wasps have evolved to use venom to defend the colony against intruders. Acid phosphatase and tachykinin, which are known as allergens and neurotoxic peptides, were found with high frequencies in the venom glands of solitary wasps. This suggests that solitary wasps might use their venom for catching and preserving prey. In the venom glands of bumblebees, little or no transcripts of major allergens or pain producing factors were identified, implying that bumblebees venoms are relatively less toxic than those of social or solitary wasps. Taken together, the differential expression patterns of venom genes in some Aculeate bees/wasps implies that bees/wasps have unique groups of highly expressed venom components, which appear to have evolved in response to both ecological and behavioral influences.

Mosquitoes are primary medical insect pests due to their diseases transmission as vectors. In Korea, the insecticide-resistant populations of disease vector mosquito species, such as Anopheles sinensis, Culex pipiens and Culex tritaeniorhynchus, have constantly increased. Thus, management of insecticide resistance to major insecticides including pyrethroids and organophosphates is required for more efficient control of resistant populations. In this study, the quantitative sequencing (QS) protocols were established to detect the frequencies of three mutations (the L1014F on voltage sensitive sodium channel and the G119S and F331W on acetylcholinesterase 1) that are associated with either pyrethroids or organophosphates. Based on the QS protocol using newly designed non-polymorphic primers, resistance allele frequencies (RAFs) were estimated in field populations of An. sinensis, Cx. pipiens and Cx tritaeniorhynchus collected from an identical site in Korea. The dynamics of each resistance allele frequency over time in the same populations were also evaluated.

The purpose of this paper was to investigate the comparison of balance and muscle strength between dominant and non-dominant legs in adults. Thirty adults in their 20s participated in this study. The dominant and non-dominant legs were selected based on the dominant hands of the target. The subject's muscle strength of legs was measured with Nicholas MMT, and the balance was measured with BIO-Rescue. We compared the dominant and non-dominant legs based on the results. The result, indicated no statistical difference on balance and muscle strength between dominant and non-dominant legs(p>.05). The results of this study will be helpful in setting the effective treatment direction and treatment level, and in controlling posture, balance and motor function.

Generally, in vivo, primary oocytes are grown and matured into secondary oocytes in the ovarian follicles. Quality of the oocytes matured in vivo is higher than that of oocytes matured in vitro, indicating the importance of materializing the microenvironment of ovarian follicles for production of high quality oocyte. Therefore, we tried to mimic the stiffness of ovarian follicles using an agarose as a biocompatible natural polymer. Unfortunately, to date, there are no many reports on whether the quality of porcine oocytes can be increased effectively under the soft matrix. Accordingly, we tried to evaluate the effects of IVM using different mechanical properties of agarose substrate on developmental competence of porcine oocytes. Agarose substrate was constructed and cumulus-oocyte-complexes (COCs) retrieved from porcine medium antral follicles were matured on non-coated (control) culture dish or dishes coated with 1% and 2% (w/v) agarose substrate. Then, cumulus expansion, embryonic development after parthenogenetic activation, and gene expression level were analyzed and compared. As the results, significant increase in blastocyst formation and cumulus expansion were detected in COCs matured on 1% (w/v) agarose substrate compared with control. Moreover, oocytes of COCs matured on 1% (w/v) agarose substrate showed significantly higher BMP15 expression level compared with control. Pro-apoptotic gene BAX expression was significantly increased in oocytes of COCs matured on 2% (w/v) agarose substrate compared with control. In the glycolytic enzyme phosphofructokinase (PFKP) gene expression, cumulus cells of COCs matured on agarose substrate showed significantly higher PFKP expression than control while they showed significantly lower BAX expression than control. These results demonstrated that quality of porcine oocytes could be increased efficiently by the IVM of immature oocytes on the soft culture matrix using agarose.

This study was conducted to evaluate the effects of insulin and epidermal growth factor (EGF) in a in vitro growth (IVG) medium on oocyte growth, in vitro maturation (IVM) and embryonic development of pig oocytes derived from small antral follicles (SAF) less than 3 mm in diameter. SAF oocytes were cultured for 2 days to induce IVG in alpha-minimal essential medium supplemented with 1 mM dbcAMP and 15% (v/v) fetal bovine serum. After IVG culture, oocyte maturation was induced by culturing IVG oocytes in IVM medium for 44 h. IVM oocytes that extruded the first polar body were selected and induced for parthenogenesis (PA) by applying electric stimulus. SAF oocytes cultured under the insulin treatment showed a significantly increased (P < 0.05) nuclear maturation (73.8%) compared to those cultured with insulin and EGF (59.8%). After PA, the proportions of blastocysts based on the number of metaphase II oocytes were significantly higher (P < 0.05) in oocytes that were cultured for IVG with insulin, EGF, and insulin + EGF (32.4%, 35.2%, and 34.8%, respectively) than in control (22.9%). IVG oocytes treated with insulin showed an increased oocyte diameter (116.3 μm) compared to those treated with insulin and EGF (114.0 μm) (P < 0.05). Intra-oocyte GSH content significantly increased (1.07 pixels/oocyte) by insulin treatment during IVG compared to that of oocytes treated with insulin + EGF (0.78 pixels/oocyte). These results demonstrate that IVG culture of SAF oocytes under insulin or/and EGF treatment supports oocyte maturation and improves embryonic development to the blastocyst stage after PA in pigs.

U0126 is a highly selective inhibitor of both MEK1 and MEK2, a type of MAPK/ERK kinase. This study was conducted to evaluate the effect of U0126 treatment during in vitro maturation (IVM) on nuclear maturation, intra-oocyte glutathione content, and embryonic development after parthenogenesis (PA). U0126 (5 μM) was supplemented to IVM medium during the first 0 (control), 2, and 4 h. The basic medium used for IVM was medium-199 supplemented with 10% (v/v) porcine follicular fluid (standard), 0.6 mM cysteine, 0.91 mM pyruvate, 75 μg/ml kanamycin, and 1 μg/ml insulin. Immature pig oocytes were matured for 44 h and then oocytes reached metaphase II stage were electrically activated to induce PA. The in vitro culture medium for embryonic development was porcine zygote medium-3 containing 0.3% (w/v) fatty acid-free BSA. When immature oocytes were treated with U0126 during the first 0, 2, 4 h of IVM culture, nuclear maturation was significantly (P < 0.05) increased by the U0126 treatment for 4 h (96.2 ± 1.3%) compared to standard IVM (90.6 ± 2.1%). Cleavage of PA embryos was significantly increased by 4 h- treatment (90.6 ± 2.2%) compared to standard medium (83.9 ± 1.8%). In addition, blastocyst formation of PA embryos was significantly (P < 0.05) increased by the treatment for 4 h (55.8 ± 5.7%) compared to 2 h (38.1 ± 6.1%). The glutathione contents in IVM oocytes were not altered by the U0126 treatments for 0, 2, and 4 h (1.28 ± 0.10, 1.16 ± 0.09, and 1.10 ± 0.09, respectively). Our results demonstrated that 5 μM U0126 treatment during the first 4 h of IVM showed positive effects on nuclear maturation, cleavage, and embryonic development in pigs.

Poor embryo quality and low blastocyst formation have been major limitations in establishment of cloned embryonic stem cells and production of cloned animals through somatic cell nuclear transfer (SCNT). Aggregation of embryos is a promising method for improving developmental competence of blastocysts. The aim of this study was to improve the blastocyst formation and the quality of parthenogenetic (PA) pig embryos by the aggregation of blastomeres at the 4-cell stage that were cultured in various type of culture dishes with or without phytohemagglutinin (PHA). The PA embryos were produced by the general method of our laboratory. On Day 2 after PA, the zona pellucida of 4 cell-stage embryos were removed by treatment with 0.5% (wt/vol) pronase solution. The 3x zona-free blastomere (ZFB) were randomly distributed in each of the following treatments for aggregation. ZFB were cultured for 5 days at 39℃ in an atmosphere 5% CO2, 5% O2, and 90% N2. In Experiment 1, effect of culture dishes on the aggregation efficiency and developmental competence of PA embryos were investigated. ZFB were cultured on non-coated (control) culture dish or dishes coated with 1% (wt/vol) agarose substrate (AS) or Well of the Well in dishes coated with 1% (wt/vol) agarose substrate (WAS). The ZFB cultured in WAS showed significantly higher (P<0.05) aggregation (81.2%) than AS and control (21.6-45.5%). The mean cell number in blastocysts derived from AS and WAS (81.4-89.3 cells/blastocyst) was significantly higher (P<0.05) than that of control (63.8 cells/blastocyst). In Experiment 2, effects of 150 ug/ml PHA treatment on the aggregation efficiency and developmental competence of embryos were investigated. The ZFB cultured in AS with PHA showed a higher (P<0.05) aggregation rate (90.0%) than that in AS without PHA, control with PHA, and control (39.2%, 57.9% and 17.5%, respectively). In conclusion, aggregation of porcine ZFB treated with PHA and agarose substrate could be a useful technique for producing improving blastocyst development with increased mean cell number of blastocysts in pigs.

We report the structural, morphological and magnetic properties of the Ni70Mn30 alloy prepared by Planetary Ball Mill method. Keeping the milling time constant for 30 h, the effect of different ball milling speeds on the synthesis and magnetic properties of the samples was thoroughly investigated. A remarkable variation in the morphology and average particle size was observed with the increase in milling speed. For the samples ball milled at 200 and 300 rpm, the average particle size and hence magnetization were decreased due to the increased lattice strain, distortion and surface effects which became prominent due to the increase in the thickness of the outer magnetically dead layer. For the samples ball milled at 400, 500 and 600 rpm however, the average particle size and hence magnetization were increased. This increased magnetization was attributed to the reduced surface area to volume ratio that ultimately led to the enhanced ferromagnetic interactions. The maximum saturation magnetization (75 emu/g at 1 T applied field) observed for the sample ball milled at 600 rpm and the low value of coercivity makes this material useful as soft magnetic material.

Incursions of red imported fire ant (RIFA), Solenopsis invicta into Korea have been increasing. After a first interception of a colony of S. invicta on Gamman pier, Pusan port while intensive surveillance by Animal & Plant Quarantine Agency (APQA) in September 2017, three more RIFA colonies have been found in sea port piers of Pyeongtek, Incheon and Pusan cities. The social forms and mitochondrial DNA haplotypes of the intercepted RIFA colonies were analysed by allelic discrimination assay (peptide nucleic acid probe based RT-PCR) of Gp-9 gene and mt-DNA fragment of 768 bp, which is part of the Cytochrome oxidaseⅠ gene. The colony on Gamman pier, intercepted in Sep. 2017 was previously reported as a haplotype 5 (H5) of mitochondrial DNA and a social form of polygyne. The colony on Hutchison pier of Pusan port, intercepted in June 2018 were confirmed as a H22 haplotype and a monogyne. Those different social forms show different origins of each colonies. Those on piers of Pyeongtek and Incheon ports, also found in 2018 were confirmed samely as H22 and monogyne. However, it could be putatively assumed that those two colonies were differently introduced via different container cargoes, considering those colonies were found in container yards of distantly located different sea ports. More genetic variation analyses using diverse sets of molecular markers such as microsatellites, genome-wide single nucleotide polymorphisms, etc. in nuclear gens are being proceeded for more exact introduction routes (origins).

The hypopharyngeal gland (HPG) of the honeybee worker produces royal jelly (RJ) and has a developmental cycle closely related to the division of labor. In this study, we investigated to compare the HPG acini diameter of differently aged worker bees with high royal jelly producing colony (HRC) or less producing colony (LRC). Additionally, we also evaluated whether the fresh weight of the head is a reliable indicator of the developmental status of HPG. The HRC showed a significantly higher RJ production about two-times as compared with those of the LRC. We measured the HG-diameters on days 1, 3, 6, 9, 12, 15. The microscopic analysis revealed that the acini size of the HRC was significantly larger than the LRC. In addition, the acini diameter of HRC was 15% longer than the LRC on the first day after emerging. It was shown that the fastest development during 3 days which is preparing for nurse the brood. The HPG acini diameters increased in both colonies in a similar fashion until day 12 and then decreased. We also compared the fresh head weight of the experimental colonies, differences were similar to the development of HPG. Therefore, high royal jelly production may have a positive correlation between HPG acini size and the fresh head weight.

To identify the venom components and their expression patterns of some Aculeata bees/wasps, venom gland-specific transcriptome analysis was conducted. FPKM values were normalized with the average of the transcription level of reference gene (a-tubulin). Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, the information on venom compositions of solitary wasps obtained in this study was not sufficient to generalizae this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insuline-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more abundant in social wasp venom. In the venom gland trancsriptome of bumblebees, major allergens or pain producing factors were barely identified, implying that bumblebee venoms are relatively less toxic than those of social or solitary wasps.

The common bed bug, Cimex lectularius, possesses a cholinesterase expressed exclusively in the salivary gland (ClSChE). In this paper, we investigated the molecular structure, tissue distribution patterns, and biochemical properties of ClSChE and showed that ClSChE exists as a soluble monomeric form or a soluble dimeric form connected by a disulfide bridge. Immunohistochemical analysis confirmed that ClSChE was expressed in the epithelial cells of both the salivary gland and the duct. In addition, the secretion of monomeric ClSChE through the proboscis during feeding was detected by western blotting using a ClSChE-specific antibody. To predict the role of ClSChE injected into the tissue of an animal host, we analyzed the extent of sequestration and hydrolysis of acetylcholine (ACh)/choline (Ch) by ClSChE by ultra-performance liquid chromatography-tandem mass spectrometry. Kinetic analysis revealed that ClSChE possesses extremely low Km (high affinity to ACh) and Vmax values. These findings suggest that ClSChE functions as a sequestering enzyme specific to ACh (not to Ch) by having a very strong affinity to ACh but an extremely long turnover time.

Porcine spermatogonial stem cells (SSCs) prefer three-dimensional (3D) culture systems to 2D ones for the maintenance of self-renewal. Of the many 3D culture systems, agar-based hydrogels are candidates for supporting porcine SSC self-renewal, and there are various types of agar powder that can be used. In this study, we sought to identify an agar-based 3D hydrogel system that exhibited strong efficacy in the maintenance of porcine SSC self-renewal. First, 3D hydrogels with different mechanics were prepared with various concentrations of Bacto agar, lysogeny broth (LB) agar, and agarose powder, and the 3D hydrogel with the strongest alkaline phosphatase (AP) activity and greatest increase in colony size was identified for the different types of agar powder. Second, among the porcine SSCs cultured in the different 3D hydrogels, we analyzed the colony formation, morphology, and size; AP activity; and transcription and translation of porcine SSC-related genes, and these were compared to determine the optimal 3D hydrogel system for the maintenance of porcine SSC self-renewal. We found that 0.6% (w/v) Bacto agar-, 1% (w/v) LB agar-, and 0.2% (w/v) agarose-based 3D hydrogels showed the strongest maintenance of AP activity and the most pronounced increase in colony size in the culture of porcine SSCs. Moreover, among these hydrogels, the strongest transcription and translation of porcine SSC-related genes and largest colony size were detected in porcine SSCs cultured in the 0.2% (w/v) agarose-based 3D hydrogel, whereas there were no significant differences in colony formation and morphology. These results demonstrate that the 0.2% (w/v) agarose-based 3D hydrogel can be effectively used for the maintenance of porcine SSC self-renewal.

The relationship of in vitro starch digestibility and gel strength was investigated at various concentrations (10-30%) of rice cultivars with different amylose contents (27.9, 17.9, and 5.2%). As the rice flour concentration increased, predicted glycemic index decreased, but gel strength increased regardless of amylose contents. Gel strength correlated strongly with amylose content, whereas in vitro starch digestibility was more highly affected by rice flour concentration than by amylose contents. Moreover, the impact of degree of gelatinization on in vitro starch digestibility of high amylose rice was also examined in terms of structural features and rheological properties. The digestion rate of fully gelatinized flour was 1.7 times higher than that of native flour, while the disrupted structure with a different gelatinization degree during starch digestion was visually demonstrated through the X-ray diffraction and molecular distribution analysis. The rice flour changed from an A-type to a V-type pattern and showed difference in crystalline melting. The low molecular weight distribution increased with increasing degree of gelatinization during starch digestion. The apparent viscosity also increased with degree of gelatinization. These results demonstrated that the starch digestibility of rice was more affected by concentration than by amylose content, as well as by the degree of gelatinization due to structural difference.