The phylogenetic relationships among the Nymphalidae (Lepidoptera: Papilionoidea) have been controversial in several perspective. The present study sequenced a total of ~ 3,500 bp from cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-1 alpha (EF-1α) in 80 nymphalid species belonging to seven subfamilies (Linmenitidinae, Heliconiinae, Nymphalinae, Apaturinae, Libytheinae, Satyrinae, and Danainae), along with those of six lycaenid species as outgroups. Phylogenetic analyses via Bayesian Inference (BI) and Maximum Likelihood (ML) algorithms concordantly supported the subfamilial relationships of (((((Linmenitidinae + Heliconiinae) + (Nymphalinae + Apaturinae)) + Libytheinae) + Satyrinae) + Danainae), with high nodal support for monophyletic subfamilies and tribes. This result is largely consistent with a previous study performed with a substantially large sequence information and morphological characters, except for the position of Libytheinae that has previously been placed as the sister to all reminder of Nymphalidae.

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation of the species we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) collected from seven Korean localities. A total of 18 haplotypes (BARCA01 ~ BARCA18), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 17 sequence types (ITS2CA01 ~ ITS2CA17), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in mitochondrial gene sequence. Phylogenetically, the mitochondrial DNA has shown several haplotypes formed independent groups with substantially high node support (≥ 90%), whereas no such grouping was evidenced for ITS2, indicating different behaviors of the two molecules. Such difference may reflect a diverse dynamics of the species such as biogeographic history, mating behaviors, and also possibly different mode of inheritance of the two molecules, but requires further scrutinized examination of the dataset. In terms of population genetic perspective, overall no population subdivision was detected from both molecules, except for locality 7 (Eocheong islet) from mitochondrial DNA. As more scrutinized analysis is performed, further fruitful inference on the geographic contour of the species might be available.

Neurotrophic factors are essential to maintain and organize neurons functionally; thereby neurotrophic factor-like substances or their inducers are expected to be applied to the treatment of neurodegenerative diseases such as stroke. In the present study, we firstly examined the effects of ethanol extracts of Hericium erinaceus (HE, Yamabushitake), on nerve growth factor expression in neuronal cells. HE extract promoted NGF expression in a brain tissue. Here we assessed neuroprotective effects of HE with a transient global cerebral ischemia model. Global cerebral ischemia was induced by occluding both common carotid arteries. Treatment with HE was initiated after ischemia induction and given once a day for 7 consecutive days. Neuronal cell loss in CA1 of hippocampus was significantly decreased and the performance in the Morris water maze was significantly improved in rats administered HE. We conclude that treatment with HE attenuated learning and memory deficits, motor functional disability, and neuronal cell loss induced by global cerebral ischemia. These results suggest that HE may be a potential candidate for the treatment of vascular dementia.

The majority of strokes are caused by ischemia and result in brain tissue damage, leading to problems of the central nervous system including hemiparesis, dysfunction of language and consciousness, and dysfunction of perception. The purpose of this study was to investigate the effects of Poly(ADP-ribose) polymerase(PARP) on necrosis in neuronal cells that have undergone needle electrode electrical stimulation(NEES) prior to induction of ischemia. Ischemia was induced in male SD rats(body weight 300g) by occlusion of the common carotid artery for 5 min, after which the blood was reperfused. After induction of brain ischemia, NEES was applied to Zusanli(ST 36), at 12, 24 and 48 hours. Protein expression was investigated using immuno-reactive cells, which react to PARP antibodies in cerebral nerve cells, and Western blotting. The results were as follows: In the cerebral cortex, the number of PARP reactive cells after 24 hours significantly decreased(p<.05) in the NEES group compared to the GI group. PARP expression after 24 hours significantly decreased(p<.05) in the NEES group compared to the GI group. As a result, NEES showed the greatest effect on necrosis- related PARP immuno-reactive cells 24 hours after ischemia, indicating necrosis inhibition, blocking of neural cell death, and protection of neural cells. Based on the results of this study, NEES can be an effective method of treating dysfunction and improving function of neuronal cells in brain damage caused by ischemia.

This study is intended to examine the motor skill learning and treadmill exercise on motor performance and synaptic plasticity in the cerebellar injured rats by harmaline. Experiment groups were divided into four groups and assigned 15 rats to each group. GroupⅠ was a normal control group(induced by saline); GroupⅡ was a experimental control group(cerebellar injured by harmaline); GroupⅢ was a group of motor skill learning after cerebellar injured by harmaline; GroupⅣ was a group of treadmill exercise after cerebellar injured by harmaline. In motor performance test, the outcome of groupⅡ was significantly lower than the groupⅢ, Ⅳ(especially groupⅢ)(p<.001). In histological finding, the experimental groups were destroy of dendrities and nucleus of cerebellar neurons. GroupⅢ, Ⅳ were decreased in degeneration of cerebellar neurons(especially groupⅢ). In immunohistochemistric response of synaptophysin in cerebellar cortex, experimental groups were decreased than groupⅠ. GroupⅢ's expression of synaptophysin was more increased than groupⅡ, Ⅳ. In electron microscopy finding, the experimental groups were degenerated of Purkinje cell. These result suggest that improved motor performance by motor skill learning after harmaline induced is associated with dynamically altered expression of synaptophysin in cerebellar cortex and that is related with synaptic plasticity.

Attacin is a well-studied glycine-rich antibacterial protein in insect immune response, which has limitary antibacterial effect to some Gram-negative bacteria. A cDNA encoding the attacin gene was screened and isolated from the immunized larvae of the swallowtail butterfly, Papilio xuthus. The complete P. xuthus attacin cDNA is 949 nucleotides encoding a 250 amino acid precursor that contains a putative 18-residue signal peptide, a common 42-residue propeptide sequence and a presumed 190-residue mature protein with a theoretical mass of 19904.01 and a pI of 9.13. The putative mature protein of P. xuthus attacin showed 48%~52% and 24%~30% identity in amino acid sequences with that of lepidopteran and dipteran insects, respectively. The attacin transcript was induced at significant level after injection with bacterial lipopolysaccharide (LPS). Recombinant attacin was highly expressed in E. coli BL21 (DE3) cells by fusing with an N-terminal S-tag/thrombin cleavage site configuration protein to avoid the cell death during induction. The expressed fusion protein was purified by Ni-NTA immobilized metal affinity chromatography (IMAC). After desalting and cleavage with thrombin, the recombinant attacin was released and showed considerably antibacterial activity against Gram-negative bacteria, E. coli ML 35. Our results proved that this protein family with a potent antibacterial activity may play a role in the immune response of butterflies.

The silkworm (Bombyx mori), as an industrial insect, possesses a high economic value. Casual discrimination and accumulated genetic information of silkworm varieties are essential ground for the practical utilization and long-term conservation. In this study, nine available microsatellite loci were successfully genotyped from ~50 silkworm strains preserved in Korea. According to genotyping analysis, we obtained 3 ~ 16 alleles per locus, with an average of 7.4, the observed heterozygosity ranging from 0.04 to 0.98, and the polymorphic information content (PIC) ranging from 0.06 to 0.88, revealing that some loci are highly variable. Among 54 strains 13 strains were casually identified by the presence of 17 strain-specific apomorphic alleles. Furthermore, 30 among remaining strains contained strain-specific allele combinations that are also apomorphic to each strain, allowing us to discriminate each of these from other strains by genotyping of multiple loci. These results collectively suggest that the silkworm microsatellite DNA is actually and potentially important molecular marker for the discrimination of the silkworm strains that are preserved as hundreds in Korea, as more loci are genotyped.

This study was performed to investigate the effects of Needle Electrode Electrical Stimulation(NEES) on ischemia-induced cere˗ brovascular accidents. After obstruction and reperfusion of arteries in white mice, the amounts of necrosis and inflammation related sub˗ stances Bax, IL-6, Caspase-3, and COX-2 were measured in neurons of the fore-brain. The following results were obtained. This study used 21 male specific pathogen free(SPF) SD rats, 8 weeks of age and approximately 300g in weight. Each exposed artery was completely occluded with non-absorbent suture thread and kept in that state for 5 minutes. The sutures were then removed to allow reperfusion of blood. Test group is control group(common carotid artery occlusion models), a GI(underwent common carotid artery occlusion), and NEES(underwent NEES after artery occlusion). The GI and NEES groups were given 12, 24, or 48 hours of reperfusion before NEES. NEES device(PG6, ITO, Japan, 9V, current, 2Hz) was used to stimulate the bilateral acupoint ST36 of the SD rats for 30 minutes while they were sedated with 3% isoflurane. An immuno-his˗ tochemistry test was done on the forebrains of the GI induced rats. Both Bax and Caspase-3 immuno-reactive cells, related to apoptosis, were greater in the GI than the NEES group. Cox-2 and IL-6 immuno-reactive cells, related to inflammation, were greater in the GI and NEES groups than the control group. We can expect that applying NEES after ischemic CVA is effective for preventing brain cells from being destroyed. And we can conclude NEES should be applyed on early stage of ischemic CVA.

The purpose of this study is to investigate the effects of group exer˗ cise program on depression in the elders with dementia. Subjects of this research were selected from the patients of Hyoja hospital(Yong˗ in, Korea). They were elders with dementia having minor to moderate degrees of cognitive function. Twenty-four subjects were randomly assigned into two groups, each with 12 people; exercise group and the control group. The control group only performed general physio˗ therapy, while the exercise group performed exercise program and general physiotherapy for 8 weeks. Depression of the exercise group and the control group were measured at baseline before the study, at 4 weeks, and at 8 weeks after the study. Depression was measured by Geriatric Depression Scale Short Form Korea Version(GDSSF-K). Firstly, when comparing the levels of depression within the same group, depression was significantly lower in the exercise group, after the group exercise program. However, depression of the control group did not show significant changes before or after the study. Secondly, when comparing the levels of depression of the control group and the exercise group, differences between the levels meas˗ ured before the study and 8 weeks after the study, was a statistically significant decrease of depression. Thirdly, effects of group exercise program according to the applied period were analyzed showing 8 weeks of group exercise to be more beneficial than 4 weeks. People who performed continuous group exercise program showed decrease in depression compared to the absence of group exercise program. This can infer beneficial effects of group exercise program. Group exercise program had desirable influence on decreasing the level of depression.

Iron catalyzes the production of free radicals, which can be related to a variety of pathological events, such as cancer and aging. The effect of dietary iron was investigated on formation of colonic aberrant crypt foci (ACF) induced by azoxymethane in male F344 rats. Animals were fed three different diets, including iron-deficient (3 ppm Fe), iron-normal (35 ppm Fe), and iron-overloaded (350 ppm Fe) diets for eight weeks. During the first and second weeks of the experiment, animals received two subcutaneous injections of azoxymethane (AOM, 15 mg/kg body weight) to induce ACF. After staining with methylene blue, the total numbers of ACF and aberrant crypts (AC) were counted on the colonic mucosa. Analysis of blood and serum was performed using a blood cell differential counter and an automatic serum analyzer. Iron-deficient diets induced a significant decrease in red blood cell counts and the values of hemoglobin concentration, hematocrit, mean corpuscular hemoglobin, and mean corpuscular volume, while an iron-overloaded diet did not affect these values. The iron-overloaded diet induced an increase in deposits of iron in the liver of rats, as determined by the ICP method and Perl’s staining. The numbers of ACF per colon showed a slight increase in iron-overloaded or iron-deficient rats, without statistical significance, compared to iron-normal rats. The number of total AC per colon in iron-overloaded rats was significantly higher than that in iron-normal rats (p<0.05). The number of large ACF (≥ 4 AC per ACF) in iron-overloaded rats was also significantly higher than that in iron-normal rats (p<0.05). These results suggest that dietary iron intake may play an important role in colon carcinogenesis in humans and animals.

Field research was undertaken to determine how barley + legume mixtures influence nitrogen fixation and N transfer to barley. Barley + legume (main plot) and 4 different sowing rate was the experiment design. Isotope dilution was used to estimate N₂ fixation and subsequent transfer to barley. Botanical composition of legume was from 1.6% to 13.9% when harvest. Legume N derived from atmosphere ranged from minus value to 58percentage in whole plant N. N transfer amount to barley varied from 12.11㎏ to 7㎏ by different method.

This study was carried out to investigate effect of herbal extracts mixtures on hair growth in an alopecia model of C57BL/6 mice. There were 4 experimental groups including distilled water (DW, negative control), 25% ethanol(EtOH, vehicle control) and two herbal extract mixtures (HE-1 and HE-2). HE-1 included Polygonum and Brassica campestris extracts and HE-2 included Mulberry root and Gardenia extracts. The 6 weeks-old C57BL/6 male mice were shaved with an electric clipper and the test compounds were topically treated with 0.2 ml per mouse daily for 3 weeks. The hair re-growth was photographically determined at days 0, 4, 7, 10, 14, 17, and 21. The number of mast cells which is an important modulator of hair growth was counted in 1 cm of dorsal skin section of mice. There were no clinical signs in all experimental groups. As the results of photometric analysis, the topical application of the herbal extracts (HE-1 & 2) for 2 weeks to dorsal skin accelerated hair re-growth remarkably faster than that of DW group (p＜0.05). Activity of alkaline phosphatase (ALP) was significantly increased in EtOH group compared to DW group (p＜0.05). Both herbal extract mixtures also increased the ALP activity, but it was not significantly different from DW. Treatment of mice with HE-2 significantly increased mast cell population compared to EtOH. Taken together, these results suggest that herbal extract mixtures used in this study may have hair-growth promoting activities and can be useful for treatment for male pattern baldness or alopecia in humans.

Bumblebees are important pollinators in greenhouse and have colonized all parts of the World. In Korea, the value of bumblebees is increasing as pollinator. However, the more recent use of reared colonies may ultimately allow pathogens to spread to peripheral areas for bumblebees. Generally, bumblebees are hosts to a large number of parasites which are mites, flies, protozoa, fungi, virus and bacteria. For most of these, very little is known about their effects against host insect, epidemiology or evolutionary ecology. So, we report pathogenic bacteria isolated from Bombus terrestris and B. ignitus at first time in Korea. Bacillus fusiformis and Klebsiella oxytoca are isolated from B. terrestris and confirmed with 16S rRNA gene nucleotide comparison in NCBI genebank. Pathogenicity of B. fusiformis was 35~40% against B. terrestris. Pantoea dispersa and K. oxytoca are isolated from B. ignitus and confirmed with 16S rRNA gene nucleotide comparison in NCBI genebank. Pathogenicity of these species were 35~40% against B. ignitus. These pathogenicity are considered as low-level.