SERPINB3 (also known Squamous cell carcinoma antigen 1, SCCA1) is involved in apoptosis, immune response, cell migration and invasiveness of cells. It has been investigated in various types of squamous cell carcinoma. Therefore we investigated the functional role of SERPINB3 gene in human epithelial ovarian cancer (EOC) using laying hens, the most relevant animal model. In 136 laying hens, EOC was found in 10 (7.4%). We compared the expression and localization of SERPINB3 using RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemistry, and SERPINB3 activation was detected in chicken and human ovarian cancer cell lines using immunofluorescence microscopy. Thereafter, we examined the prognostic value of SERPINB3 expression in patients with EOC by multivariate linear logistic regression and Cox’ proportional hazard analyses. In present study, SERPINB3 mRNA was induced in cancerous ovaries (p< 0.01), and it was only expressed in the glandular epithelium(GE) of cancerous ovaries of laying hens. SERPINB3 protein was localized predominantly to the nucleus of glandular epithelium in cancerous ovaries of laying hens, and it was abundant in the nucleus of both chicken and human ovarian cancer cell lines. In 109 human patients with EOC, 15 (13.8%), 66 (60.6%) and 28 (25.7%) of those patients showed weak, moderate and strong expression of SERPINB3 protein, respectively. Strong expression of SERPINB3 protein was a prognostic factor for platinum resistance (adjusted OR, 5.94; 95% Confidence Limits, 1.21-29.15). Therefore SERPINB3 may play an important role in ovarian carcinogenesis and be a novel biomarker for predicting platinum resistance and a poor prognosis for survival in patients with EOC. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010- 0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

Acute ischemic stroke results from sudden decrease or loss of blood supply to an area of the brain, resulting in a coinciding loss of neurological function. The antioxidant action of melatonin is an important mechanism among its known effects to protective activity during ischemic/reperfusion injury. The focus of this research, therapeutic efficacy of melatonin on recovery of neurological function following long term treatment in ischemic brain injured rats. Male Sprague-Dawley rats (n=40; 8 weeks old) were divided into the control group, and MCAo groups (Vehicle, MT7 : MCAo+ melatonin injection at 7:00, MT19 : MCAo+melatonin injection at 19:00, and MT7,19 : MCAo+melatonin injection at 7:00 and 19:00). Rat body weight and neurological function were measured every week for 8 weeks. After 8 weeks, the rats were anesthetized with a mixture of zoletil (40 mg/kg) and xylazine (10 mg/kg) and sacrificed for further analysis. Tissues were then collected for RNA isolation from brain tissue. Also, brain tissues were analyzed by histological procedures. We elucidated that melatonin was not toxic in vital organs. MT7,19 was the most rapidly got back to mild symptom on test of neurological parameter. Also, exogenous melatonin induces both the down-regulation of detrimental genes, such as NOSs and the up-regulation of beneficial gene, including BDNF during long term administration after focal cerebral ischemia. Melatonin treatment reduced the loss of primary motor cortex. Therefore, we suggest that melatonin could be act as prophylactic as well as therapeutic agent for neurorehabilitative intervention.

This study was carried out to investigate characteristic pattern of fruiting body of Ganoderma lucidum and their antioxidant activity. Mycelia of all strains were firstly inoculated into potato dextrose agar(PDA) and then transfered to a media of saw dust which contained 20% rice bran. These mycelia of saw dust were then inoculated into oak tree in polyethylene bags which has been sterilized for 8h at 120℃. The polyethylene bags were sent to a growth room for growth of fruit bodies. Antioxidant activities of each fruiting body were investigated by DPPH method.

The EAG (Electroantennogram) has been used to find out chemical substance effective to insect. We tried to use EAG recording for catching of responsible acoustic signals to Lirimyza trifolii (Diptera: Agromizidae). In fly auditory organ known as Johnston's organ located on antenna. Pure tone sounds (sine wave) and courtship songs of several kinds of species were broadcasted to excised head of the fly. Significant EAG signals were detected when released the 20 Hz pure tone sound and long pulse train patterns, which were courtship songs of Cryptonevra inquilina and Lipara japonica (Chloropidae). For conformation of it's effectiveness, we further analyzed movement behavior with the signal got from the EAG recording. Information entropy could represent the specimens activities of movement tracks. The entropy with sound was relatively lower than without sound. The combination of EAG responding and behavioral observation would be useful to analyze the response of insect pest against acoustic signals and develop acoustic pest managing tool in agriculture.

In vivo redox reaction is involved in the processes of oxidative diseases. Thus, direct and non-invasive measurement of in vivo free radical reactions in living animals should be essential to understanding the roles of free radicals in pathophysiological phenomena. Electron spin resonance (ESR) technique has been utilized in analysis of free radicals, which are generated through imbalance of in vivo redox status. In vivo ESR/spin probe technique using nitroxide radicals as spin probes was developed for estimation of in vivo free radical reactions in whole living animals. This technique using a probe may become a powerful tool for use in clarifying mechanisms of disease and in monitoring pharmaceutical therapy. The application of ESR was introduced and discussed in this article.

Melatonin is induced by light information through the retina and leads to growth factor activation. Thus, we investigated the effects of melatonin by controlling the photoperiod of growing young rats. Male Sprague‐gDawley rats (n=6; 4 weeks old) were divided into two experimental groups: the L/D group (normal photoperiod; light/dark: 12/12 h; lights on at 9:00 a.m.) and the L/L group (light/light: 24 h). Rat body weight and food consumption were measured daily for 8 weeks. After 8 weeks, the rats were anesthetized with a mixture of ketamine (50 mg/kg) and xylazine (10 mg/kg) and sacrificed. Tissue was then collected for RNA isolation (from brain, heart, liver, kidney, adrenal gland, testis, tibia, hind limb muscles). Also, serum was isolated from blood using a centrifugal separation. The L/L group had significantly lower body weight than the L/D group from 4 to 6 weeks (p<0.05). The L/D group had increased tissue mass, compared with the L/L group, but the difference was not statistically significant. The L/D group had a significantly higher melatonin concentration than the L/L group between the hours of midnight and 2:00 a.m (p<0.01). These results indicate that photoperiod length may affect the secretion of melatonin from the pineal gland. Also, the reduction of nocturnal melatonin secretion may retard the development of growing young rats. In future studies, we plan to compare exogenous melatonin administration with endogenous melatonin concentration induced by photoperiod control. Moreover, we will confirm whether the effects seen in pathological animal models can be reversed by controlling the photoperiod.

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.

Previous studies indicated that Matsumuraeses phaseoli and M. falcana (Lepidoptera: Tortricidae) are separate species since a few differences were observed in genitalia morphology and female sex pheromone composition. A clear difference was detected in the DNA sequences of cytochrome oxidase I of the two species separately collected in different plants and regions. A hybridization test also showed that a post-zygotic reproductive isolation occurred between the species. In field monitoring, however, both species have been caught simultaneously and together in the separate sex pheromone traps installed for the two species around neighboring soybean and red bean fields. Molecular marker-assisted identification with several adults sampled from the trapped insects showed that only ca. 40% of M. phaseoli adults identified as the species by genitalia morphology was the M. phaseoli, while ca. 97% of M. falcana adults identified as the species was the M. falcana. The result indicated that the observation of genitalia did not make a decisive criterion for classification of the insects. Conclusively, it suggested that the sex pheromones of the two species should be studied more precisely although there is a possibility that the two species are hybridized in fields as in laboratory, and speciation is under process.

Pregnancy is a unique event in which a fetus develops in the uterus despite being genetically and immunologically different from the mother, and the underlying mechanisms remain poorly understood. To analyze the differential gene expression profiles in nonpregnant and 7 days post coitus (dpc) pregnant uterus of mice, we performed a global proteomic study by 2‐D gel electrophoresis (2‐DE) and MALDI‐TOF‐MS. The uterine proteins were separated using 2‐DE. Approximately 1,000 spots were detected on staining with Coomassie brilliant blue. An image analysis using Melanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between pregnant and nonpregnant uterus. Twenty‐one spots were identified as differentially expressed proteins, of which 10 were up‐regulated proteins such as alpha‐fetoprotein, chloride intracellular channel 1, transgelin, heat‐shock protein beta‐1, and carbonic anhydrase II, while 11 were down‐regulated proteins such as X‐box binding protein, glutathione S‐transferase omega 1, olfactory receptor Olfr204, and metalloproteinase‐disintegrin domain containing protein TECADAM. Most of the identified proteins appeared to be related with catabolism, cell growth, metabolism, regulation, cell protection, protein repair, or protection. Our results uncovered key proteins of mouse uterus involved in pregnancy.

Matsumuraeses phaseoli and M. falcana cannot be classified with the external morphological characters. Although differences in the morphology of male genitalia and in the mitochondrial DNA sequence of cytochrome oxidase I between the two species have been found, there is no information that the two species are biologically different species till now. We, therefore in this study, tried to cross the two species to observe postzygotic incompatibility in the next hybrid generations and to know whether the two species are 'biological species' or not. In crossing between the parents, two kinds of F1 hybrids were produced successfully. In inter- and intra-breeding between F1 hybrids, two lines crossed with females of F1 hybrid produced by females of M. phaseoli could not produce F2 adults to show inviability of larvae. The other two lines produced F2 adults successfully and the F2 adults produced F3 larvae in inbreeding. In back-crossing between parents and F1 hybrids, the two lines of 8 breeding lines, in which females of F1 hybrid were produced by females of M. phaseoli, could not produce the next generation of larvae. The other six lines produced F2 adults successfully. The results indicated that maternal factors of F1 hybrid produced by M. phaseoli females contributed to create the incompatibility between the F1 females and other lines. In conclusion, the results showed a postzygotic reproductive isolation between M. phaseoli and M. falcana in part.

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.

(E,E)-8,10-dodecadienyl acetate (E8E10-12:Ac) and (E)-8-dodecaenyl acetate (E8-12:Ac) have been selected as the candidate chemicals for sex pheromone components of the M. phaseoli, female through GC-EAD tests, whereas the two compounds and an additional candidate, (E,Z)-7,9-dodecadienyl acetate (E7Z9-12:Ac), have been found at a ratio of 7:1:1 in the abdominal tip extract (Yum et al., 2008). In order to determine the actual composition of sex pheromone, therefore, several blends using the three chemicals were evaluated for attractiveness to males of M. phaseoli around red bean and soybean fields. Individual components as well as two blends consisted of E8E10-12:Ac/E7Z9-12:Ac and E8-12:Ac/E7Z9-12:Ac did not show attractiveness, whereas the blend of E8E10-12:Ac/E8-12:Ac showed an increased effect in male capture. Of the tested blends with all three chemicals, the 7:1:2 composition of E8E10-12:Ac, E8-12:Ac and E7Z9-12:Ac attracted the most number of males. The results suggested that E7Z9-12:Ac is one of the sex pheromone components and may act as a synergist.

The techniques of IVM, IVF and IVC of canine oocytes may provide useful information for gamete salvage programs and the conservation of endangered canidae. This investigation has been made to determine the efficiency of in vitro maturation (IVM) as a basic experiment to study the development of canine oocytes after in vitro fertilization (IVF). The rate of oocytes developing to the MII stage was higher in the hormone treated group (10 IU/ml hCG+eCG, 14.7%, p<0.05) than in the control group (0 IU/ml hCG+eCG, 10.0%). The monospermy and pronuclear rates of canine oocytes were investigated after caffeine treatment on IVF. Canine oocytes were fertilized in the Fert‐TALP medium supplemented with 0, 10, 20 or 30 mM caffeine (Fert I, Fert II, Fert III or Fert IV, respectively). The highest pronuclear formation rate was obtained in the Fert I for 24 h IVF (6.7%, 6/89). Therefore, it is believed that unlike in other mammals, caffeine in canine IVF does not increase the efficiency of fertilization rate, and is not an important factor.

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.

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.