Climate change effects are particularly apparent in many cool-season grasslands in South Korea. Moreover, the probability of climate extremes has intensified and is expected to increase further. In this study, we performed climate change vulnerability assessments in cool-season grasslands based on the analytic hierarchy process method to contribute toward effective decision-making to help reduce grassland damage caused by climate change and extreme weather conditions. In the analytic hierarchy process analysis, vulnerability was found to be influenced in the order of climate exposure (0.575), adaptive capacity (0.283), and sensitivity (0.141). The climate exposure rating value was low in Jeju-do Province and high in Daegu (0.36–0.39) and Incheon (0.33–0.5). The adaptive capacity index showed that grassland compatibility (0.616) is more important than other indicators. The adaptation index of Jeollanam-do Province was higher than that of other regions and relatively low in Gangwon-do Province. In terms of sensitivity, grassland area and unused grassland area were found to affect sensitivity the most with index values of 0.487 and 0.513, respectively. The grassland area rating value was low in Jeju-do and Gangwon-do Province, which had large grassland areas. In terms of vulnerability, that of Jeju-do Province was lower and of Gyeongsangbuk-do Province higher than of other regions. These results suggest that integrating the three aspects of vulnerability (climate exposure, sensitivity, and adaptive capacity) may offer comprehensive and spatially explicit adaptation plans to reduce the impacts of climate change on the cool-season grasslands of South Korea.

This study aimed to examine the effect of a mild elevation in serum cholesterol level in a porcine coronary overstretch restenosis model using a balloon angioplasty catheter or drug-eluting coronary stent. Pigs were divided into two groups and were fed a commercial normal diet (CND, n = 4) or a high-fat diet (HFD, n = 4) for 5 weeks. Coronary overstretch injury by balloon angioplasty or stent implantation was induced in the left anterior descending and left circumflex artery after 1 week of feeding. Histopathological analysis was performed at 4 weeks after coronary injury. During the experiment, the total cholesterol level in the HFD group increased by approximately 44.9% (from 65.9 ± 3.21 mg/dL at baseline to 95.5 ± 9.94 mg/dL at 5 weeks). The lumen area in the CND group was reduced in comparison with that in the HFD group after balloon angioplasty. After stent implantation, the injury score showed no significant difference. There were significant differences in the neointimal area (2.7 ± 0.33 mm2 in the CND group vs. 3.3 ± 0.34 mm2 in the HFD group, p<0.05), lumen area (2.6 ± 0.54 mm2 in the CND group vs. 2.0 ± 0.33 mm2 in the HFD group, p<0.05), and percent area stenosis (52.0 ± 7.96% in the CND group vs. 62.4 ± 5.15% in the HFD group, p<0.05). Body weight change was not different between the two groups. Increased serum cholesterol level activated vascular smooth muscle cell proliferation in the porcine coronary overstretch model.

The mosquito is a dangerous disease vector, which transfers various diseases like dengue virus, Zika virus and malaria.This study used an alternative way of mosquito repellent agents rather than chemical repellents such as DEET. We haveemployed a novel emitters of ultra-sonic sounds, which showed repellent effects to mosquito behaviors that avoid ultra-sonicsounds. Using NGS analysis after ultrasonic treatment, we found that some subsets of sensory and other related geneswere affected, indicating that ultrasonic sounds affects gene expression associated with host finding behaviors in Aedesaegypti. Additionally, we performed electrophysiological recording and behavior assay testing repellency in a chamberin Culex pipiens, which is the most common species in Korea.

We performed a survey for flavivirus infection and distribution of Aedes albopictus that known as Zika and Dengue virus vector using black–light trap and BG-sentinel trap around urban area in Korea. Mosquitoes were collected in 27 cities during March to November (twice a month) year 2016. Total numbers of mosquitoes collected 102,102 including 19 species 8 genera during collecting period. Total 21,467 Ae. albopictus was collected that 20,961(24.3%) by BG-sentinel trap and 506 (3.2%) by Black-light trap in urban area. Trap index(trap/night) of Ae. albopictus was showed highest in Hamyang (TI:992.3) and lowest in Taebaek (TI:0.3) there was only collected by Black-light trap. A total of 894 pools from all collecting Ae. albopictus were performed a Flavivirus detection. Flavivirus was not detected during study period. This study may provide basic information for surveillance of imported diseases (include Zika virus) and vectors in Korea.

Taste substances are recognized by gustatory sensory neurons that express putative seven transmembrane proteins in the gustatory receptor (Gr) family. However, the gustatory tuning of the molecular receptors encoded by these gustatory receptor genes remains unknown in honey bees. Here we first functionally characterize a gustatory receptor responding to umami taste L-amino acids in the western honey bee, Apis mellifera. Using Ca2+ imaging assay and two-voltage clamp recording, we first report that one of the gustatory receptors of honeybee, AmGr10, functions as a selectively tuned amino acid receptor in taste neurons. In addition, we report a floating electrode-based bioelectronic tongue mimicking honeybee taste systems for the detection and discrimination of umami substances. This floating electrode-based bioelectronic tongue mimicking insect taste systems can be a powerful platform for various applications such as food screening, and it also can provide valuable insights on insect taste systems.

One of the overlooked points in mosquito blood feeding research is a final step before blood feeding. We provide the anatomical and chemosensory evidence that a piercing structure of the mouthpart of the mosquitoes is an essential apparatus for the penultimate stage in blood feeding in mosquitoes. Indeed, the stylet possesses a number of sensory hairs located at the tip of the stylet. These hairs house olfactory receptor neurons that express two conventional olfactory receptors of Aedes aegypti (AaOrs), AaOr8 and AaOr49, together with the odorant co-receptor (AaOrco). In vivo calcium imaging using transfected cell lines demonstrated that AaOr8 and AaOr49 were activated by volatile compounds present in blood. Taken together, we identified olfactory receptor neurons in the stylet involved in mosquito blood feeding behaviors, which in turn indicates that olfactory perception in the stylet is necessary and sufficient for mosquitoes to find host blood in order to rapidly acquire blood meals from a host animal.

This study was conducted to evaluate the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To determine the movement of degradable HGC nanoparticles in the body, 20 mg/kg of lutetium177-labeled HGC (Lu177-HGC) with the size ranging from 320 to 400 nm was injected intravenously into ICR mice, and the amount of radioactivity remaining in blood and several organs was measured at various time points during the period of 5 days. In the pharmacokinetics analysis using the Lu177 radioisotope, the free Lu177 was mainly distributed and accumulated in the order of kidney>liver>lung at 1 day after the injection of the radioisotope. However, the Lu177-HGC showed a high distribution of nanoparticles in the order of liver>spleen>kidney during the experimental period of 5 days. These results would provide a basic pharmacokinetics for the use of HGC as a drug carrier in drug delivery system.

The ultimate goal of this study is to assess the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To understand the movement of degradable nanoparticle HGC in the body, we intravenously injected a dose of 20 mg/kg of Cy5.5-labeled HGC with size ranging from 320 to 400 nm into ICR mice, and measured the amount of fluorescence remaining in blood and several organs at various time intervals. In blood, the level of Cy5.5-labeled HGC was the highest at 15 min, then after 30 min it decreased rapidly and reached a plateau form 30 min to 28 days. In the tissue we confirmed the presence of nanoparticles at high levels in the order of kidney>liver>submandibular gland until 28 days after injection. However, we did not find the presence of the particles in the brain or testes. These results will provide basic information on HGC as a drug delivery agent.

Although it is believed that internal nutrient sensors play important roles in feeding behaviors, their molecular and neural mechanisms underlying of the modulation of physiological status and cell growth are poorly understood. Using a Ca2+ imaging experiments with heterologous expression systems, we show that one of the gustatory receptors in the western honey bee Apis mellifera is selectively tuned to amino acids. Remarkably, we report that this gustatory receptor of the honey bee is highly expressed in hypopharyngeal gland, which plays a role in caste differentiation as well as royal jelly production and secretion. Knocking down this gustatory receptor gene reduces cellular pathways responsible for nutritional sensing such as mTOR signals in hypopharageal gland. Furthermore, the interfering expression of this gustatory receptor gene not only alters morphological changes and developmental retardation of the hypopharyngeal gland, but it also blocks cellular growth signals to induce autophagy. This new report indicates that internal sensing and downstream signals detecting nutrients is essential for honey bee to maintain the cellular growth and development of internal organs essential for caste development and maintenance of social structure in the honey bee.

Insect pollinators of the endanger orchid Cypripedium japonicum were surveyed and identified during two years, as a part of a conservation project of the orchid at Jukyeup-san and Hwaak-san (Mt.), South Korea. In total 40 individuals of 16 species in 4 families were identified. The dominant family was Halictidae, and Lasioglossum exiliceps Vachal visited the most frequently C. japonicum during the surveys. The average visiting frequency was 2.5 individuals per hour and the highest 4.3, from 12:00 – 13:00 in a day. After 15:00 insects did not visit the flowers at all. However, all of the visiting insects were found to not carry a pollinium or pollens of the orchard on their bodies; pollen carryover by any of the visiting insects did not occur at all. The orchid seems to require certain pollinators in particular body thickness due to its unique pollination mechanism. The orchid has two exit route openings, around 1 cm in diametre, where the entrapped insects can exit and an anther is situated just in front of each opening. It was inferred that a pollen carrier should be around 1 cm in body thickness. Therefore, the candidate species as the proper pollen carriers can be Tetralonia nipponensis Perez, Xylocopa appendiculata circumvolans Smith and Bombus consobrinus Dahlbom among the surveyed visitors.

Olfactory sensitivity exhibits daily fluctuations. Several studies have suggested that the olfactory system in insects is modulated by both biogenic amines and neuropeptides. However, molecular and neural mechanisms underlying olfactory modulation in the periphery remain unclear since neuronal circuits regulating olfactory sensitivity have not been identified. Here, we investigated the structure and function of these signaling pathways in the peripheral olfactory system of the American cockroach, Periplaneta americana, utilizing in situ hybridization, qRT-PCR, and electrophysiological approaches. We showed that tachykinin was co-localized with the octopamine receptor in antennal neurons located near the antennal nerves. In addition, the tachykinin receptor was found to be expressed in most of the olfactory receptor neurons in antennae. Functionally, the effects direct injection of tachykinin peptides, dsRNAs of tachykinin, tachykinin receptors, and octopamine receptors provided further support for the view that both octopamine and tachykinin modulate olfactory sensitivity. Taken together, these findings demonstrated that octopamine and tachykinin in antennal neurons are olfactory regulators in the periphery. We propose here the hypothesis that octopamine released from neurons in the brain regulates the release of tachykinin from the octopamine receptor neurons in antennae, which in turn modulates the olfactory sensitivity of olfactory receptor neurons, which house tachykinin receptors.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

The yellow fever mosquito, Aedes aegypti, is a vector for transmitting dengue fever and yellow fever. An assessment was made of the histopathological and molecular effects of pellitorine, an isobutylamide alkaloid, on third instar Ae. aegypti larvae. At 5 mg/L concentration of pellitorine, whole body of the treated larvae became dark in color, particularly damaged thorax and abdominal regions. Pellitorine targeted mainly on midgut epithelium and anal gills, indicating variably dramatic degenerative responses of the midgut through a sequential epithelial disorganization. The anterior and posterior midgut was entirely necrosed, bearing only gut lumen residues inside the peritrophic membranes. Pellitorine caused comprehensive damage of anal gill cells and branches of tracheole and the debris was found in hemolymph of anal gills. RT-PCR analysis indicates that the compound inhibited gene expression encoding V-type H+-ATPase and aquaporine 4 after treatment with 2.21 mg/L pellitorine. The results provide a fact that pellitorine merits further study as a potential larvicide with a specific target site or a lead molecule for the control of mosquito populations.

The present study purposed to examine the effects of transcutaneous electrical nerve stimulation, self-stretching and functional massage on the recovery of muscle contraction force for muscle fatigue caused by sustained isotonic contraction. The subjects of this study were 45 healthy students. They were divided into transcutaneous electrical nerve stimulation group(n=15), self-stretching group(n=15) and functional massage group(n=15), and using Primus RS. We observed the pattern of changes in maximal voluntary isometric contraction force(MVIC) after causing muscle fatigue in quadriceps femoris muscle through sustained isotonic contraction. Maximal voluntary isometric contraction force(MVIC) were greatly increased after transcutaneous electrical nerve stimulation, self-stretching and functional massage. In the comparison of recovery rate of muscle contraction force for muscle fatigue caused by sustained isotonic contraction among the treatment groups, it did not show any significant differences. However, it showed that each treatment may be effective in recovery of muscle fatigue caused by sustained isotonic contraction.

We investigated the molecular and kinetic properties of two acetylcholinesterases (AmAChE1 and AmAChE2) from the Western honey bee, Apis mellifera. Western blot analysis revealed that AmAChE2 has most of catalytic activity rather than AmAChE1, further suggesting that AmAChE2 is responsible for synaptic transmission in A. mellifera, in contrast to most other insects. AmAChE2 was predominately expressed in the ganglia and head containing the central nervous system (CNS), while AmAChE1 was abundantly observed not only in the CNS but also in the peripheral nervous system/non-neuronal tissues. Both AmAChEs exist as homodimers; the monomers are covalently connected via a disulfide bond under native conditions. However, AmAChE2 was associated with the cell membrane via the glycophosphatidylinositol anchor, while AmAChE1 was present as a soluble form. The two AmAChEs were functionally expressed with a baculovirus system. Kinetic analysis revealed that AmAChE2 has approximately 2,500-fold greater catalytic efficiency toward acetylthiocholine and butyrylthiocholine than AmAChE1, supporting the synaptic function of AmAChE2. In addition, AmAChE2 likely serves as the main target of the organophosphate (OP) and carbamate (CB) insecticides as judged by the lower IC50 values against AmAChE2 than against AmAChE1. When OP and CB insecticides were pre-incubated with a mixture of AmAChE1 and AmAChE2, asignificant reduction in the inhibition of AmAChE2 was observed, suggesting a protective role of AmAChE1 against xenobiotics. Taken together, based on their tissue distribution pattern, molecular and kinetic properties, AmAChE2 plays a major role in synaptic transmission, while AmAChE1 has non-neuronal functions, including chemical defense.

Alzheimer’s disease (AD) is a fatal disorder wherein patients suffer from sensory, motor, and cognitive loss. Currently, the identification and validation of biomarkers for diagnosing AD and other forms of dementia are increasingly important. Olfactory dysfunction is present in patients diagnosed with Alzheimer’s disease or idiopathic Parkinson's disease. Alzheimer’s patients show neuropathological changes in areas of the brain central to the olfactory processing center, suggesting the theoretical importance and potential diagnostic utility of investigating functional changes in olfaction in these patients. However, the usefulness of olfactory screens to serve as informative indicators of Alzheimer’s is precluded by the lack of knowledge regarding neural and molecular mechanisms underlying olfactory dysfunction onto Alzheimer's diseases. To test these ultimate questions, we used molecular and electrophysiological recording techniques to find out the difference of olfactory responses and AD related protein expression patterns by using fly model, Drosophila melanogaster that over-expresses the human β -amyloid, tau protein. We postulated that such flies would present with progressive olfactory impairments compared with age-matched wild type control flies. In this study, our hypothesis is that there is a correlation between olfactory deficits and the spatial expression pattern of β-amyloid and tau protein deposition. Therefore, we demonstrate a specific concentration of lesions in central olfactory structures such as antenna and Maxillary palps. Our study indicates that deficits on olfactory identification may occur in AD, which will be valuable as an indicator of neuropathogenesis.

Cinnamaldehyde as the main component of Cinnamomum plants is well known as mammalian transient receptor potential ankyrin 1 (TRPA1) agonist, also activated by low temperature stimuli and mechanosensation. The other TRP subfamily, transient receptor potential vanilloid-1 (TRPV1) sensitive to pungent compounds such as capsaicin and allicin mediates the feeling of warmth. Both TRPA1 and TRPV1 channels are abundantly distributed in sensory neurons. Thus, there is possibility that these channels modulate repellent behaviors of mosquitoes and Drosophila through olfactory receptor neurons (ORNs). In order to confirm this hypothesis, we carried out laboratory repellent tests with cinnamaldehyde to Aedes aegypti females using arm-in-cage test and to a wild type and two TRP channel mutants Drosophila lines using a choice assay. Cinnamaldehyde showed strong repellency against Ae. aegypti and Drosophila wild adults at tested concentrations. However, a mutant fly line did not discriminate or detect the existence of the repellent. These behavioral data suggest that cinnamaldehyde may directly target the TRP channel. More studies to elucidate neural correlates of repellency to ainnamaldehyde compound are as follows: 1) Identifying the ORNs mediating cinnamaldehyde detection using single-sensillum recording techniques, 2) Co-localization of TRP genes on olfactory organs of Ae. aegypti and Drosophila using in situ hybridization and 3) Whether the Aedes TRP homologs might function in cinnamaldehyde repellency using rescued TRP chennels of Drosophila.