The objective of this experiment was to investigate the effect of drip irrigation volume on tomatoes (Solanum lycopersicum L.) grown in a greenhouse using perlite medium. Plants were treated by three different irrigation treatment I0, I25, and I50 (where irrigation volume of I25 and I50 was 25% and 50% higher than I0, having limited or no leaching). Growth characteristics of plants, yield and water use efficiency were measured. The result showed that plant height, leaf length and leaf width were lowest in the I0 treated plants. However, these parameters were not statistically significant differences between the plants that were grown in the I25 and I50 treatment. Soluble solids content, acidity and dry matter of 111th, 132nd, and 143rd days harvested tomato were higher in the plants irrigated with lowest volume (I0) than the higher volume (I25 or I50). In addition, water content was lower in the 111th and 132nd days of harvested tomatoes from the I0 treatment. The number of big-size tomatoes (>180 g) was significantly higher in the I25 irrigated plants. There was no significant difference in the total number of harvested fruits among the treatments. The average fruit weight and total yield of harvested tomatoes were lowest in the I0 treated plants. The water consumption of tomato was not significantly different amongst the treatments but water use efficiency was lowest in the I0 treatment. Principal component analysis revealed that total soluble solid and acidity of tomato showed a positive correlation between each other. These results suggest that I25 was the optimum irrigation treatment for tomato based on its measured growth characteristics, yield and water use efficiency.

Feral cats are widely considered to be leading the potential impacts on public health. This study aimed to provide estimates of vital data for feral cats relating Trap-Neuter-Return (TNR) to establish strategies effectively to manage feral cats in Pyeongtaek. Thus, this study focused on estimating feral cat population in Pyeongtaek and conducted a comparative analysis of the data for feral cats in Seoul (2013). The number of feral cats was estimated from 23,069 to 26,655 in Pyeongtaek, 2019. In relation to human population, when comparing the number of feral cats of Pyeongtaek and Seoul, it ranged from 4.57% to 5.28%, and from 1.97% to 2.55% respectively. This showed that Pyeongtaek was higher than Seoul. Fewer kittens were found in high-density areas, which the TNR project is believed to be generally effective in controlling the number of feral cats. In conclusion, in urban and rural complexes such as Pyeongtaek City, the number of feral cats compared to the population was higher than that of Seoul City, and the TNR program is believed to be somewhat effective in controlling the number of feral cats. When implementing TNR, it is necessary periodically to investigate the population and reflect them in policymaking.

Calcium is the most abundant stored mineral in the human body and is especially vital for bone health; thus, calcium deficiency can cause bone-related diseases, such as osteopenia and osteoporosis. However, a high concentration of serum calcium, which is commonly known as hypercalcemia, can also lead to weakened bones and, in severe cases, osteosarcoma. Therefore, it is necessary to maintain the concentration of calcium that is appropriate for bone biology. In the present study, we aimed to elucidate the effects of high concentration of calcium, approximately 2 folds the normal calcium level, on osteoblast differentiation. The CaCl2 treatment showed dose-dependent suppression of the alkaline phosphatase activity and mineralized nodule formation. Calcium showed cytotoxicity at an extremely high concentration, but a moderately high concentration of calcium that results in inhibitory effects to osteoblast differentiation showed no signs of cytotoxicity. We also confirmed that the CaCl2 treatment repressed the mRNA expression and protein abundance of various osteogenic genes and transcriptional factors. Considered together, these results indicate that a high concentration of calcium negatively regulates the osteoblast differentiation of C2C12 cells.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

Autophagy is an important self-eating process to eliminate damaged or unused organelles. We identified nine autophagy-related genes (Atg) including AaAtg-1, -3, -4b, -4d, -5, -6, -8, -12 and -13 from the Asian tiger mosquito, Aedes albopictus. Developmental expression patterns indicate that mRNA levels of AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 were highly expressed in egg, whereas expression of AaAtg8 was high in 1stand3rdinstarlarvalstages. TissuespecificexpressionofthesegenesindicatesthatAaAtg1 was highly expressed in thorax and midgut in blood-fed adult female mosquitoes (BF), and head and thorax in sugar fed adult female mosquitoes (SF). Transcript level of AaAtg3 was high in thorax in BF, but head, thorax and Malpighian tubules in SF. AaAtg4b, -4d mRNA levels were significantly high in Malpighian tubules in BF, and head in SF, respectively. AaAtg-5 and -6 transcripts were highly expressed in head in BF, and expression of AaAtg-8 was high in Malpighian tubules in BF. Levels of AaAtg-12 and -13 mRNAs were significantly high in head and midgut in BF. Induction patterns of AaAtg genes against pathogens showed that AaAtg-1, -3, -4b, -8, -12 and -13 were strongly induced at 6 h-post injection of S. aureus, and mRNA levels of AaAtg-1, -3 and -13 were significantly induced by E. coli challenge after 3 h-post injection in SF abdominal carcass. In SF midgut, AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 transcripts were drastically induced at 9 h-injection of E. coli and S. aureus, while expression of AaAtg-8 was highly induced by S. aureus and C. albicans at 9 h-post injection. Each AaAtg gene was slightly induced by E. coli, S. aureus or C. albicans at different time points in abdominal carcass in BF. Interestingly, AaAtg-8 was not induced by microbial challenge. While eight other Atg genes except AaAtg-8 were highly influenced by S. aureus at 6 and 9 h-post injection, E. coli at 3 h-post-treatment, and 3, 6, and 9 h-post inoculation. In the future, we will characterize the functional roles of autophagy during mosquito-microbes interaction.

It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

Host defense against pathogen invasion highly relies on immune defense machinery that is controlled by the nuclear factor-κB (NF-κB) of transcription factors. The Toll pathway are well known as an insect innate immune mechanism to protect host itself from invaded pathogens. Basically, in the edible insect, Tenebrio molitor, the Toll pathway is primarily activated by polymeric Lys-type peptidoglycans (PGNs), and components of fungal cell walls, β-1,3-glucan. Based on the current studies, the tremendous study has been focused on recognition and subsequent activation of spätzle in haemolymph, hence, there is a grave gap for intracellular event. Herein, in order to understand intracellular event of Toll signaling pathway, the Dorsal gene were identified. Moreover, domain analyses of TmDorsal2 gene indicate that there are two major domains such as Rel homology domain (RHD), ig-like, plexins, and transcription factors (IPT) domains. Based on the achieved results, TmDorsal2 mRNA was highly expressed in 1-day old pupa. Furthermore, TmDorsal2 was highly expressed in Malpighian tubules and fat body in last instar larvae (LL), and likewise mainly expressed in Malpighian tubules during adult 5-day old period, also the lowest expression of TmDorsal2 was observed in gonads. Moreover, TmDorsal2 mRNA levels after infection with E. coli appreciably went up at 6 and 9h time points. To investigate the effects of TmDorsal2 RNAi on larval susceptibility against various pathogens namely E. coli, S. aureus or C.albicans, dsRNA of TmDorsal2 has been synthesized the larvae dissected after 24h. As a result, TmAttacin1a, 1b and 2, TmDefencine1 and 2, TmTenecin1, 2, 3 and 4, TmCecropin2, TmColeoptericin1 and 2, Thaumatin-like protein 1 and 2 markedly reduced in the gut after injecting all mentioned microbes. In contrast, TmTenecin 2, Thaumatin-like protein 1 and 2 strikingly increased after microbe injection in the fat body. Interestingly, the most AMPs gene expression in whole body experimental case were upregulated. On the horizon, we will investigate effects of TmDorsal1 RNAi on larval susceptibility against various pathogens. Taken together, our studies may aid to understand insect innate immunity.

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

Micropropagation studies of two Hydrangea. macrophylla cultivars, ‘Blaumeise’ and ‘Vanilla Sky’, were performed by tissue culture. The results showed that supplementing B5 medium with different concentrations of 6-Benzylaminopurine (BA) (0.0, 0.5, 1.0, 1.5, and 2.0 mgL-1) significantly increased the number of shoots and leaves per explant compared with the control in both cultivars. In ‘Blaumeise’, 1.0 mgL-1 of BA was the concentration that achieved the highest number of shoots and leaves, i.e., 12.5 and 93.25, respectively. In ‘Vanilla Sky’ 1.5 mgL-1 of BA resulted in the highest numbers of shoots (4.25) and leaves (29). The longest shoot in both ‘Blaumeise’ (2.10 cm) and ‘Vanilla Sky’ (1.18 cm) was obtained at 0.0 mg·L-1 of BA. The highest number of roots in both ‘Blaumeise’ (11) and ‘Vanilla Sky’ (6.5) were obtained with indole-3-butyric acid (IBA) 1.0 mgL-1 of IBA.

To be better fit for highways, pavement systems are required to provide comfortable and safe driving and be structurally durable. Composite pavements can be an effective option as they are more durable by placing a high functional asphalt overlay on a rigid concrete base layer. In order to apply a composite pavement system to the field, it is necessary not only to develop technologies that prevent reflecting crack and deterioration of the base layer, but also to improve bonding performance of materials and ensure structural performance as a pavement system against traffic loading. In advanced countries like Japan, USA and Europe, high-functional composite pavement systems are being put into practice across new highway networks. In this study, we evaluated structural performance (rutting, reflecting crack, and deflection) by applying traffic loads of actual highways through an accelerated pavement tester (APT) of a composite pavement section made up of a quiet porous surface laid over a water-proofing layer, a continuously reinforced concrete base, and a lean concrete sub-base layer, which was developed with new pavement methods used for each layer prior to field application. The APT specimen was constructed with paving materials and equipment actually used on site in the same dimensions (W3.5m*L14m*H2m) as actual highway sections in Korea, and 3-axle double-wheel heavy load (45ton) cart type KALES(Korean Accelerated Loading and Environmental Simulator) traveling on the specimen in both directions was used to simulate traffic loading. After applying around 8,574,000 ESALs of traffic loads, no reflecting crack occurred on the asphalt surface of the composite pavement, without surface distress except for rutting. In order to examine what causes rutting of pavements, we surveyed thickness of pavements by layer and measured asphalt density.