Apolipophorin-III (apoLp-III) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insects. Recently, apolipophorin-III in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of the apoLp-III in insects. To investigate the genes which have a relationship with apoLp-III in fall webworm larvae, we reduced endogenous Hc apoLp-III mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-III reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-III, which induced an elevated level of superoxide anion in Hyphantria cunea larvae. The observed effect of Hc apoLp-III RNAi suggests that Hc apoLp-III is related to the action/expression of antioxidants, especially MnSOD.
A new insect member of the STAT family of transcription factors (HcSTAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and transcribed in hemocyte, fat body, midgut, epidermis, and Malpighian tubule. Especially, hemocyte and Malpighian tubule showed transcriptional activation of HcSTAT upon Gram-negative and -positive bacteria challenge. Gram-negative and -positive bacteria challenge specifically results in nuclear translocation of HcSTAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vivo treatment with sodium orthovanadatetranslocates HcSTAT to the nucleus in hemocyte cells.
Effects of Vegemil® containing soybean proteins and isoflavones on the growth and bone density of broiler chickens were investigated. One-week-old male and female Arbor Acres broiler chickens were fed on Vegemil® A containing 3% soybean proteins and 162 ppm isoflavones, instead of water, for 30 days and their growth indices (body weight, leg weight and femur length) and bone density were analyzed. The body weight gains in male and female chickens were increased by 15.6% and 31.7%, respectively, following feeding Vegemil® A compared to normal water. Vegemil® A increased leg weight as well as femur length of females by 22.9% and 15.0%, respectively. In addition, Vegemil® A feeding enhanced femoral bone density by 21.3% in comparison with water feeding. Therefore, it is suggested that Vegemil® A not only facilitates body growth, but also strengthens bone density of normal chickens, and that it could be a promising candidate for the improvement of infant growth and for the prevention of menopausal osteoporosis.
Reactive oxygen species (ROS) is toxic to living organisms, because its high reactivity causes oxidative damage to proteins, nucleic acids, and lipids. Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on the cloning of MnSOD cDNA from Hyphantria cunea and its induction upon bacterial infection and various stresses. The open reading frame of MnSOD is composed of 645 bp, encoding 215 amino acid residues. The theoretical molecular mass and pI of putative MnSOD was evaluated to be 24276 Da and 9.14, respectively. The MnSOD from H. cunea is highly similar to human MnSOD (59.5%) as well as Bombyx mori MnSOD (76.2%). MnSOD showed no big induction upon bacterial infection and stresses, compared to that of Cu/ZnSOD.
Apolipophorin-Ⅲ (apoLp-Ⅲ) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insect. Recently, apolipophorin-Ⅲ in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of apoLp-Ⅲ in insects. To investigate the genes which have a relationship with apoLp-Ⅲ in fall webworm larvae, we reduction of endogenous Hc apoLp-Ⅲ mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-Ⅲ reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-Ⅲ, which induced an elevated level of superoxide anion in H. cunea larvae. The observed effect of Hc apoLp-Ⅲ RNAi suggests that Hc apoLp-Ⅲ is related to the action/expression of antioxidants.
Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.
Reactive oxygen species (ROS) is toxic to living organisms, because its high reactivity causes oxidative damage to proteins, nucleic acids, and lipids. Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on the cloning of MnSOD cDNA from Hyphantria cuneaand its induction upon bacterial infection and various stresses. The open reading frame of MnSOD is composed of 645 bp, encoding 215 amino acid residues. The theoretical molecular mass and pI of putative MnSOD was evaluated to be 24276 Da and 9.14, respectively. The MnSOD from H. cunea is highly similar to human MnSOD (59.5%) as well as Bombyx mori MnSOD (76.2%). MnSOD showed no big induction upon bacterial infection and stresses, compared to that of Cu/ZnSOD.
Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.
Background : Perilla frutescens L. is valuable as a medicinal plant as well as a natural medicine and functional food. Limonene perilla collected from various places showed 60% limonene compounds. However biological activity of these accession has not been reported before. Therefore, this study was conducted to investigate the biological activity of limonene perilla. Methods and Results : Fractional solvent extracts were obtained by using organic solvents such as n-hexane, chloroform, ethyl acetate, n-BuOH, and aqueous solvent from different parts of limonene perilla extracted initially in 70% EtOH. We investigated the effects of limonene perilla on total phenol and flavonoid contents, FRAP (Ferric Reducing Antioxidant Power), total saponin contents and tyrosinase inhibition activity. Leaves of limonene perilla produced the highest total phenolic contents (29.88 mg·CAE/g), flavonoid (8.39 mg·QE/g) and saponin contents (47.77 mg·GIE/g) than stems and roots of limonene perilla. FRAP of leaves was 823.00±3.58 μM·FeSO4·E/mg. Tyrosinase inhibition activity rate was 40.31% in 70% ethanol extracts from leaves of limonene perilla. Conclusion : This results suggest that leaf of limonene perilla fractions has significant antioxidant activity. Also, limonene perilla could be used as a functional biomaterial in developing cosmetics and functional foods.
Background : Plants are the rich source of antioxidants, which plays a very important role in maintaining human health. Their antioxidant property protects cells of different organs of human beings against free radicals and free radical mediated diseases. Even though, there is lack of knowledge on the antioxidant effect of lutein present in plants. In the present study, lutein was isolated from the GreenTea leaves (Camellia sinensis) which is used as a dietary source. Methods and Results : The procedure adopted for the isolation and purification of lutein using acetone extraction and preparative high performance liquid chromatography (HPLC) is simple and less time consuming. Free radicals scavenging activity of isolated lutein from acetone extract of GreenTea was assessed by DPPH radical scavenging assay and reducing power. The isolated lutein scavenged 79% of DPPH radicals at 20 ㎍/㎖ and two fold lower concentration compared to the standard antioxidants (α-tocopherol). No significant differences were found between the reducing power of the lutein and BHT when their concentrations were high. However, significant differences were observed at relatively low concentrations, the reducing power of lutein was isolated from the GreenTea leaves was stronger than those of their acetone extract and standard antioxidants (BHA). Both electron spin resonance (ESR) and in vitro assay confirmed that lutein was isolated from the GreenTea leaves, exhibited a greater capacity for scavenging superoxide (O2 •-) and hydroxyl (OH •) radicals than standard antioxidants β-carotene and α-tocopherol respectively. Conclusion : The results proven that lutein isolated from GreenTea leaves has an efficient antioxidant ability, it could serve as an antioxidant to scavenge reactive oxygen species.
Background : Lutein, a xanthophyll, consists of chains with 8 conjugated double bounds containing closed rings on each end of the chain. This carotenoid is found in fruits and vegetables, especially dark green leafy vegetables such as green tea. In this study, we investigated the anticancer effects of purified lutein from green tea on human cancer cell lines containing prostate carcinoma cancer cells (LNCaP). Methods and Results : Prostate carcinoma cancer cells (LNCaP) were cultured and evaluated the inhibitory effect of lutein isolated from green tea compared other carotenoids (β-carotene and lycopene) on cell proliferation. Cyclin D1 and PCNA were evaluated as cell differentiation. In results, PCNA/cyclin regulates the initiation of cell proliferation by mediating DNA polymerase. Under cultural conditions, lycopene remarkably suppressed the PCNA expression prostate cancer cell line LNCaP in higher doses (20 μM - 100 μM) statistically. However, β-carotene and lutein presented the less inhibitory effects on PCNA expression. Determination of PCNA expression in control and treated cells demonstrates that lycopene did affect proliferation in LNCaP cancer cells in dose-dependent manner. However, β-carotene and lutein suppressed the cyclin D1 expression in dose-dependent manner but no in lycopene group. These results indicate that differ carotenoids presented the various suppressive ability of PCNA and cyclin D1 expression in cell proliferation. Conclusion : In conclusion, lutein suppressed the carcinogenesis of induced prostate cancer cell line by acting as a suppressor for inhibiting the expression of cyclin D.