To clarify the anti-obesity effect of Allomyrina dichotoma larvae (ADL), we previously reported that ADL block adipocyte differentiation on 3T3-L1 cell lines through downregulation of transcription factors, such as peroxisome proliferator-activated receptor-γ (PPARG) and CCAAT/enhancer binding protein-α (CEBPA). In this study, we tested whether ADL prevent obesity in mice fed a high-fat diet (HFD) and further investigated the mechanism underlying the effects of ADL. All mice were maintained on a normal-fat diet (NFD) for 1 week and then assigned to one of five treatment groups: (1) NFD; (2) HFD; (3) HFD and 100 mg·kg−1·day−1 ADL; (4) HFD and 3000 mg·kg−1·day−1ADL; or (5) HFD and 3000 mg·kg−1·day−1 yerba mate (Ilex paraguariensis, positive control). ADL and yerba mate were administered orally daily. Mice were fed experimental diets and body weight was monitored weekly for 6 weeks. Our results indicated that ADL reduced body weight gain, organ weight and adipose tissue volume in a dose-dependent manner. Body weight gain was approximately 22.4% lower compared to mice fed only HFD, but the difference did not reach the level of statistical significance. Real-time polymerase chain reaction (PCR) analysis revealed that gene expression levels of PPARG, CEBPA and lipoprotein lipase (LPL) in the epididymal fat tissue of HFD-fed mice receiving 3000 mg·kg−1·day−1 ADL were reduced by 12.4-, 25.7-, and 12.3-fold, respectively, compared to mice fed HFD only. Moreover, mice administered ADL had lower serum levels of triglycerides and leptin than HFD-fed mice that did not receive ADL. Taken together our results suggest that ADL and its constituent bioactive compounds hold potential for the treatment and prevention of obesity.

Although Korean horn beetle (Allomyrina dichotoma) larvae have been long used as an oriental medicine for the treatment of liver diseases in Korea, there is little scientific evidence of their effects. In this study, we investigated the anti‐obesity activity of A. dichotoma larvae extract (ADE) by using 3T3‐L1 cells as an in vitro model of adipogenesis. To analyze its role as an inhibitor of adipogenesis, 3T3‐L1 cells were treated with both ADE and differentiation inducer (DM) at the same time, and then the amount of lipid was quantified. We then analyzed the mRNA and protein expression level of adipogenic or lipogenic genes. The results showed that formation of lipid droplets in differentiated 3T3‐L1 cells was markedly decreased, and triglyceride content was reduced by 22.15% in response to 2000 μg/mL ADE. In addition, ADE downregulated mRNA and protein expression of transcription factors peroxisome proliferator‐activated receptor (PPAR)‐γ and CCAAT/enhancer binding protein (C/EBP)‐α implicated in adipogenesis, and significantly decreased the expression levels of adipocyte fatty acid binding protein (aP2), fatty acid synthase (FAS), stearoyl‐coenzyme desaturase‐1 (SCD1), and lipoprotein lipase (LPL) related to lipogenesis. Our results demonstrate that ADE suppressed adipogenesis and lipogenesis in 3T3‐L1 cells, suggesting that ADE has anti‐obesity activity as a food supplement.

Recently, the Tenebrio molitor larva was recognized as a novel food ingredient by the Ministry of Food and Drug Safety in Korea. Accordingly, we investigated its physical and sensory characteristics to establish the cooking conditions that may increase the demand of T. molitor larvae as a food. In this study, T. molitor larvae were cooked by various methods such as hot air dry, oven-broil, roast, pan fry, deep fry, boil, steam, and by microwave. In the physical evaluation of texture, the hardness and fracturability values were highest when larvae were cooked in the microwave. The adhesiveness, springiness, and chewiness values were highest when larvae were boiled. Boiled and steamed larvae had the highest lightness (L value), while oven-broiled larvae had the highest redness (a value) and yellowness (b value) values. Sensory evaluations assessed the appearance, aroma, flavor, and texture of cooked T. molitor larvae. Steamed and boiled larvae sizes were significantly large and the form was well preserved similar to fresh larvae. The moisture heat cooked (steamed and boiled) T. molitor larvae had the aroma and flavor of steamed corn, canned pupa, and boiled mushroom. In case of oven-broiled T. molitor larvae, the aroma and flavor of mealworm oil, seafood, sweet and roasted sesame were higher than in those cooked by other methods. In texture among sensory evaluation, the hardness and crispiness were the highest in the hot air dried and oven-broiled larvae, whereas juiciness was significantly higher in the boiled and steamed. Accordingly, we suggest that oven-broiled T. molitor larva will be prefered by consumer, due to its the rich aroma and flavor.

To examine the expression profile of oxidative stress responsive genes in Spodoptera litura, we constructed a cDNA library from S. litura injected with hydrogen peroxide (H(2)O(2)). Using a microarray chip composed of 2,964 cDNAs, we screened gene expression at 1, 3, 5, 7, and 9 h post H(2)O(2) injection. Data were clustered into 15 groups of genes that behave similarly across each time course. Seventy-three genes were identified as being at least twofold up- or downregulated after treatment with H(2)O(2) in S. litura. We constructed expressed sequence tags (ESTs) for genes that changed at least twofold after treatment with H(2)O(2) . The functional classification of these ESTs based on Gene Ontology showed that the ESTs are rich in genes involved in oxidoreductase activity (5.7%), defense (14.3%), cellular process (22.9%), and development (17.1%).

The baculovirus expression vector system (BEVS) is one of the powerful insect cell systems for heterologous protein expression. However, over-expression of heterologous proteins in this system sometimes results in protein misfolding and aggregation because of insufficient levels of folding catalysts. In previous study using the differential screening (DS) method, we isolated only 40 differentially expressed genes after treatment with tunicamycin, an unfolded protein response (UPR) inducer. To isolate more protein folding catalysts from insect, we performed suppressive subtractive hybridization (SSH) with untreated and tunicamycin-treated Bm5 cell lines in this study. We could isolate 366 differentially expressed clones by SSH method and produced expressed sequence tags (ESTs). ESTs included the UPR pathway-related genes involved in protein folding, including heat shock proteins, molecular chaperones, foldases, as well as glycosylation and secretory pathway related genes. Identification of the tunicamycin responsive genes using SSH provides more information about the UPR-related genes in insect cells, and will facilitate modifications of the protein folding pathway in the ER to improve heterologous protein expression.

Drosophila melanogaster has been used as a useful model to study development and disease. In this study, we established the primary culture method of Drosophila in the intestine to understand how intestinal stem cells (ISCs) mediate tissue repair during infection and disease. To obtain intestinal cells, we separated intestines from adult flies and isolated single cells by enzymatic treatment. The survival of cultured cells was measured using MTS-analysis. The maximum growth rate of the cells was observed on the 9th day after seeding. In addition, the presence of ISCs and enteroendocrine cells was confirmed by delta and prospero staining. Accordingly, we supposed that Drosophila melanogaster gut cells established in this study are probably useful in studies about intestinal stem cell regulation and various diseases occurring in the intestine.

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