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.

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0～10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500～600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro‐matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM‐199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1 kV/cm for 30 μs in 0.3 M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium‐3 (PZM‐3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM‐3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine‐porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat‐porcine and porcine‐bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.

Human embryonic stem (ES) cells retain the capacity for self‐renewal, are pluripotent and differentiate into the three embryonic germ layer cells. The regulatory transcription factors Oct4, Nanog and Sox2 play an important role in maintaining the pluripotency of human ES cells. The aim of this research was to identify unknown genes upregulated in human ES cells along with Oct4, Nanog, and Sox2. This study characterizes an unknown gene, named chromosome 1 open reading frame 31 (C1orf31) mapping to chromosome 1q42.2. The product of C1orf31 is the hypothetical protein LOC388753 having a cytochrome c oxidase subunit VIb (COX6b) motif. In order to compare expression levels of C1orf‐ 31 in human ES cells, human embryoid body cells, vascular angiogenic progenitor cells (VAPCs), cord‐blood endothelial progenitor cells (CB‐EPCs) and somatic cell lines, we performed RT‐PCR analysis. Interestingly, C1orf31 was highly expressed in human ES cells, cancer cell lines and SV40‐immortalized cells. It has a similar expression pattern to the Oct4 gene in human ES cells and cancer cells. Also, the expression level of C1orf31 was shown to be upregulated in the S phase and early G2 phase of synchronized HeLa cells, leading us to purpose that it may be involved in the S/G2 transition process. For these reasons, we assume that C1orf31 may play a role in on differentiation of human ES cells and carcinogenesis.

The principal objective of this study was to assess the antioxidative activities of Petasites japonicus against oxidative stress in bovine brain tissue . Petasites japonicus is found with a relatively widespread distribution, and is cultivated as a culinary vegetable in Korea. Petasites japonicus samples were dried either by freeze-drying or by hot air-convection drying (80℃), then evaluated for their antioxidative activity by measuring 1-dipheny-1，2-picrylhydrazyl (DPPH) radical scavenging, and by measuring thiobarbituric acid-reactive substances (TBARS) in brain homogenates subjected to Fe2+ -mediated lipids with or without the addition of botanical extract. Hot air convection-drying resulted in a slight increase in the extraction yie1d as compared with freeze-drying. However, total phenol and flavonoid contents in freeze-dried Petasites japonicas were significantly higher than those of hot air convection-drying. Freeze-drying increased the free radical scavenging activity of Petasites japonicas, leaves, and stems by 52.6, 28.6, and 248.0%, as compared with hot air convection-drying. Additionally, the IC50 values measured by TBARS in hot air convection-dried Petasites japonicas， leaves, and stems were increased by 36.0, 31.6, and 15.9%, as compared to those of freeze-drying. Although antioxidative activity was reduced slightly by heat processing in Petasites japonicas, freeze-drying for each portion of Petasites japonicus was the most appropriate for use as a functional food and pharmaceutical material.

As indigenous aphid parasitoid, Aphelinus varipes kill aphids for feeding in addition to parasitization. Because of this characteristic of A. varipes, this parasitoid may have the possibility of biological control agent against aphids. So we have evaluated traits such as daily paratization, total parasization, number of aphids killed by host feeding, sex ratio, development time, pupal mortality of A. varipes parasitizing green peach aphid, Myzus persicae. At 25°C and 16L:8D, longevity, total paratization and host feeding of A. varipes female was 11.0, 25.3, and 63.3 days, respectively. And development time of male and female, sex ratio (M:F), pupa mortality of offspring of A. varipes were 12.0 days, 12.5 days, 0.88, and 11.6%, respectively. However, because these results are not enough to estimate potential of A. varipes as biological control agents/factors, other factors such as host suitability (Macrosiphum euphorbiae, Aulacorthum solani), effect of temperature, and host seeking behavior of A. varipes continually will be investigated.

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.

As an effective generalist predator of aphids and other hemipteran pests, Harmonia axyridis has been a successful biological control agent. Interestingly, it was known that there were varied in color patterns on H. axyridis elytra. In fact, Seo & Youn (2007) reported that H. axyridis had five color patterns, for example, succinea 1, 2, conspicua, spectabilis, and axyridis. But there are uncertain that H. axyridis elytra colour patterns are regulated by genetic polymorphism. So we tried to what is the reason that color patterns are greatly variable. To identify DNA markers linked to a elytra polymorphism, amplified fragment length polymorphism (AFLP) analysis was performed on DNA samples from four female succinea, conspicua, spectabilis and Coccinella septempunctata which is another species in Coccinellidae. AFLP analysis with the restriction endonuclease combination EcoRⅠ and MseⅠwas performed. Using 12 AFLP primer pairs, nine AFLP fragments which is specific between succinea, conspicua, spectabilis was identified. These nine AFLP fragments were isolated, cloned and sequenced. Subsequent UPGMA cluster analysis revealed three major group of H. axyridis populations. These genetic tree showed that H. axyridis elytra colour diversity was affected by genetic polymorphism. For more genetically understanding elytra colour genes, different primer combinations may be need to be generate enough polymorphic markers. These genetic analyses may be facilitate the understanding of molecular mechanism behind wing colour pattern formation.