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Anti-inflammatory effects of glycosaminoglycan (GAG) derived from cricket (Gryllus bimaculatus, Gb) were investigated in a complete Freund’s adjuvant (CFA)-treated chronic arthritic rat model. This GAG produced a significant anti-edema effect as evidenced by inhibition of C-reactive protein (CRP) and rheumatoid factor, and interfered with atherogenesis by reducing proinflammatory cytokine levels of (1) vascular endothelial growth factor (VEGF) production in human umbilical vein endothelial cells (HUVEC), (2) interleukin-6, (3) prostaglandin E2-stimulated lipopolysaccharide in RAW 264.7 cells, and (4) tumor necrosis factor (TNF)-α production in normal splenocytes, in a dose-dependent manner. This GAG was also found to induce nitric oxide (NO) production in HUVEC cells and elevated endothelial nitric oxide synthase (eNOS) activity levels. Histological findings demonstrated the fifth lumbar vertebrae (LV) dorsal root ganglion, which was linked to the paw treated with Gb GAG, was repaired against CFA-induced cartilage destruction. Further, combined indomethacin (5 mg/kg)–Gb GAG (10 mg/kg) inhibited more effectively CFA-induced paw edema at 3 h and 2 or 3 d after treatment to levels comparable to only the anti-inflammatory drug indomethacin. Ultraviolet (UV)-irritated skin inflammation also downregulated nuclear factor κB (NFκB) activity in transfected HaCaT cells. Data suggest that the anti-inflammatory effects of GAG obtained from cricket (Gb) may be useful for treatment of inflammatory diseases including chronic arthritis. [ABSTRACT FROM PUBLISHER]

Protaetia brevitarsis larvae have been used as a traditional medicine to treat various liver diseases. Freeze-dried powder and solvent partitioned fractions of Protaetia brevitarsis larvae, in this study, were examined for hepatoprotective and antitumor activities in vivo and in vitro. Oral administration of the powder reduced signs of acute and chronic liver injuries in diethylnitrosamine-induced hepatotoxic mouse model. Various cancer cell types, but not normal primary hepatocytes, were killed by the hexane, ethyl acetate and butanol fractions, which were partitioned from larval extracts. Apoptosis and autophagy were primarily induced in cancer cells by hexane and ethyl acetate fractions, respectively, and can be the cytotoxic mechanisms of the fractions. Further analyses of the insect fractions could lead to identification of novel bioactive substances that can be exploited as therapeutics for various human liver diseases. [ABSTRACT FROM AUTHOR]

Amyloid-β peptide ( Aβ) generation initiated by β-site amyloid precursor protein cleaving enzyme 1 BACE1 is a critical cause of Alzheimer's disease. In the course of our ongoing investigation of natural anti-dementia resources, the ethyl acetate ( EtOAc) fraction exerted strong BACE1-specific inhibition with the half maximal inhibitory concentration (IC50) value of 9.2 × 10−5 μg/mL. Furthermore, Aβ(25-35)-induced cell death was predominantly prevented by the EtOAc fraction of Allomyrina dichotoma larvae through diminishing of cellular oxidative stress and attenuating apoptosis by inhibiting caspase-3 activity. Taken together, the present study demonstrated that A. dichotoma larvae possess novel neuroprotective properties not only via the selective and specific inhibition of BACE1 activity but also through the alleviation of Aβ(25-35)-induced toxicity, which may raise the possibility of therapeutic application of A. dichotoma larvae for preventing and/or treating dementia. [ABSTRACT FROM AUTHOR]

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 3 T3- L1 cells as an in vitro model of adipogenesis. To analyze its role as an inhibitor of adipogenesis, 3 T3- 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 3 T3- 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 (a P2), 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 3 T3- L1 cells, suggesting that ADE has anti-obesity activity as a food supplement. [ABSTRACT FROM AUTHOR]

Abstract: The centipede Scolopendra subspinipes mutilans has been a medically important arthropod species by using it as a traditional medicine for the treatment of various diseases. In this study, we derived a novel lactoferricin B like peptide (LBLP) from the whole bodies of adult centipedes, S. s. mutilans, and investigated the antifungal effect of LBLP. LBLP exerted an antifungal and fungicidal activity without hemolysis. To investigate the antifungal mechanism of LBLP, a membrane study with propidium iodide was first conducted against Candida albicans. The result showed that LBLP caused fungal membrane permeabilization. The assays of the three dimensional flow cytometric contour plot and membrane potential further showed cell shrinkage and membrane depolarization by the membrane damage. Finally, we confirmed the membrane-active mechanism of LBLP by synthesizing model membranes, calcein and FITC-dextran loaded large unilamellar vesicles. These results showed that the antifungal effect of LBLP on membrane was due to the formation of pores with radii between 0.74nm and 1.4nm. In conclusion, this study suggests that LBLP exerts a potent antifungal activity by pore formation in the membrane, eventually leading to fungal cell death. [Copyright &y& Elsevier]

Highlights: [•] l- and d-CopA3 was designed by replacing an amino acid of CopN5 derived from coprisin. [•] l- and d-CopA3 exerted potent antifungal activity without hemolysis. [•] l- and d-CopA3 had membrane permeabilization mechanism in the fungal cells. [•] Only d-CopA3 maintained a potent antifungal activity under proteolytic condition. [•] d-CopA3 has potential as a novel antimicrobial agent. [Copyright &y& Elsevier]

The antihypertensive effects of both extracts and glycosaminoglycan derived fromIsaria sinclairii(IS) were investigated in a spontaneously hypertensive rat (SHR) model. Groups of rats were treated orally with 30 mg/kg each of: (1) saline control or extracts of (2) water-IS (3) methanol-IS, (4) butanol-IS, (5) ethyl acetate-IS, or (6) captopril as positive control. The 30-mg/kg dose was administered with a standard diet every day for a period of 2 wk. The antihypertensive effects of the individual extracts were in the following order: methanol > water > ethyl acetate > butanol. Glycosaminoglycan (GAG) obtained from IS as a water-soluble alcohol precipitation fraction produced an antihypertensive effect. One month following administration of GAG derived from IS to SHR animals there was a marked decrease in systolic blood pressure from 183 to 105 mm Hg and reduced diastolic blood pressure from 148 to 80 mm Hg compared to untreated control SHR rats. It was found that GAG produced an antihypertensive effect, which was more effective than the positive control captopril. In the SHR animal model a fall of 19% in body weight was observed in the group that received GAG. Data thus indicate that GAG derived fromI. sinclairiimay be a potent, naturally occurring antihypertensive agent. SHR (Spontaneously Hypertensive Rat); WKY (Wistar Kyoto Rat); glycosaminoglycan (GAG);I. sinclairii(IS); ISMC30:I. sinclairiimethanol extract 30 mg/kg; ISG2:I. sinclairiiglycosaminoglycan 2 mg/kg; IS GAG:I. sinclairiiglycosaminoglycan.This study was supported by the Rural Development Administration, Basic research project Pj006642 (2009–2012). [ABSTRACT FROM AUTHOR]

Coprisin is a 43-mer defensin-like peptide from the dung beetle, Copris tripartitus. In this study, we tested its minimum inhibitory concentration and performed combination assays to confirm the antibacterial susceptibility of coprisin and synergistic effects with antibiotics. The synergistic effects were evaluated by testing the effects of coprisin in combination with ampicillin, vancomycin, and chloramphenicol. The results showed that coprisin possessed antibacterial properties and had synergistic activities with the antibiotics. To understand the synergistic mechanism(s), we conducted hydroxyl radical assays. Coprisin alone and in combination with antibiotics generated hydroxyl radicals, which are highly reactive oxygen forms and the major property of bactericidal agents. Furthermore, the antibiofilm effect of coprisin alone and in combination with antibiotics was investigated. Biofilm formation is the source of many relentless and chronic bacterial infections. The results indicated that coprisin alone and in combination with antibiotics also had antibiofilm activity. Therefore, we conclude that coprisin has the potential to be used as a combinatorial therapeutic agent for the treatment of infectious diseases caused by bacteria. [ABSTRACT FROM AUTHOR]

We isolated transferrin cDNA from tobacco cutworm ( Spodoptera litura) and refer to it as SlTf ( Spodoptera litura transferrin). The 2,237-bp SlTf cDNA encoded 685 amino acids, with a predicted Mw of 76.3 kDa and an isoelectric point of pH 7.97. The amino acid sequence of the SlTf protein had 11-81% similarity with those of other reported animal transferrins, showing the highest similarity with another Lepidopteran insect, the silkworm ( Bombyx mori), and the lowest similarity with atlantic cod ( Gadus morhua) serum transferrin. Phylogenetic tree analysis showed that SlTf was close to transferrins of B. mori and M. sexta. By urea-polyacrylamide gel electrophoresis, four different iron-bound forms (apo-, C-terminal monoferric, N-terminal monoferric and diferric) were found from both SlTf and human transferrin, suggesting the C-lobe iron-binding motif of SlTf possesses the iron-biding activity, although its amino acid sequence is not well conserved compare to that of vertebrate transferrins. Accordingly, we suggest that the amino acid residues of iron-binding sites in SlTf are different from those of human serum transferrin, however the iron-binding capacity is conserved in both the C-lobe and the N-lobe of SlTf. [ABSTRACT FROM AUTHOR]

Abstract: Coprisin is a 43-mer defensin-like peptide from the dung beetle, Copris tripartitus. Here, we investigated the induction of apoptosis by coprisin in Candida albicans cells. Coprisin exerted antifungal and fungicidal activity without any hemolytic effect. Confocal microscopy indicated that coprisin accumulated in the nucleus of cells. The membrane studies, 1,6-diphenyl-1,3,5-hexatriene, calcein-leakage, and giant unilamellar vesicle assays, confirmed that coprisin did not disrupt the fungal plasma membrane at all. Moreover, the activity of coprisin was energy- and salt-dependent. Next, we investigated whether coprisin induced apoptosis in C. albicans. Annexin V-FITC staining and TUNEL assay showed that coprisin was involved with both the early and the late stages of apoptosis. Coprisin also increased the intracellular reactive oxygen species level, and hydroxyl radicals were included at high levels among the species. The effect of thiourea as a hydroxyl radical scavenger further confirmed the existence of the hydroxyl radicals. Furthermore, coprisin induced mitochondrial membrane potential dysfunction, cytochrome c release, and activation of metacaspases. In summary, this study suggests that coprisin could be a model molecule for a large family of novel antimicrobial peptides possessing apoptotic activity. [Copyright &y& Elsevier]

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. [ABSTRACT FROM AUTHOR]

The nucleotide sequences containing an entire genomic region and 5′ upstream region of Antheraea yamamai fibroin gene have been determined. The gene consists of an initial exon encoding 14 amino acids, an intron (150 bp), and a long second exon coding for 2641 amino acids. The fibroin coding sequence shows a specialized organization with a highly repetitive region flanked by non repetitive 5′ and 3′ ends. Northern blot analyses confirmed that fibroin gene is actively expressed in the posterior silk gland of the final instar larvae of Antheraea yamamai. [ABSTRACT FROM AUTHOR]