Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

Most traditional genome sequencing projects involving infectious viruses include culturing and purification of the virus. This can present difficulties as an analysis of multiple populations from multiple locations may be required to acquire sufficient amount of high-quality DNA for sequence analysis. The electrophoretic method provides a strategy whereby the genomic DNA sequences of the Korean isolate of Pieris rapae granulovirus (PiraGV-K) were analyzed by purifying it from host DNA by pulsed-field gel electrophoresis, thus simplifying sampling and labor time. The genomic DNA of infected P. rapae was embedded in agarose plugs, digested with a restriction nuclease and methylase, and pulsed-field gel electrophoresis (PFGE) was used to separate PiraGV-K DNA from the DNA of P. rapae, followed by mapping of fosmid clones of the separated viral DNA. The double-stranded circular genome of PiraGV-K encodes 120 open reading frames (ORFs), covering 92% of the sequenced genome. BLAST and ORF arrangement showed the presence of 78 homologs to other genes in the database. The mean overall amino acid identity of PiraGV-K ORFs was highest with the Chinese isolate of PiraGV (~99%), followed up with Choristoneura occidentalis ORFs at 58%. PiraGV-K ORFs were grouped, according to function, into 10 genes involved in transcription, 11 involved in replication, 25 structural protein genes, and 15 auxiliary genes. Genes for Chitinase (ORF 10) and cathepsin (ORF11), involved in the liquefaction of the host, were found in the genome. The recovery of PiraGV-K DNA genome by pulse-field electrophoretic separation from host genomic DNA had several advantages, compared with its isolation from particles harvested as virions or inclusions from the P. rapae host. We have sequenced and analyzed the 108,658 bp PiraGV-K genome purified by the pulsed field electrophoretic method. The method appears to be applicable to the analysis of genomes of large viruses. The chitinase, identified by PiraGV-K genome sequence, was functionally characterized by quantitative PCR, Western blot analysis, immunohistochemistry and transmission electron microscopy.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

This study aims to address the effect of interferential current therapy and thereby to provide basic resources to physical therapies for internal medicine by identifying symptoms for functional dyspepsia and serum gastrin level that shows gastric motility. Following results were obtained by performing interferential current therapy to 16 subjects composed of 8 for functional dyspepsia group and 8 for control group for 20 minutes a day, 3 days a week, for 6 weeks. In control group, serum gastrin level was significantly( p<.01), lowered after the therapy whereas there was no significant difference observed in all questions from questionnaire for symptoms of functional dyspepsia between before and after the therapy. In functional dyspepsia group, serum gastrin level was significantly(p<.01), lowered after the therapy and there was also significant(p<.01) reduction in every question from questionnaire for symptoms of functional dyspepsia between before and after the therapy. There was more significant decrease in serum gastrin level and reduction in questionnaire for symptoms of functional dyspepsia in the functional dyspepsia group compared to the control group(p<.01). This study confirms the interferential current therapy as an effective therapeutic method for internal diseases including functional dyspepsia since it not only improves the symptoms of functional dyspepsia but also allows the gastric motility close to normal.

The mushrooms have been used as traditional medicines and food resources in many countries. The objective of this study was to determine antioxidant compounds and to evoluate tyrosinase inhibitory activity of extracts from Hypsizigus marmoreus. The pileus and stipe of Hypsizigus marmoreus were extracted with methanol and water, separately. The methanol extract of Hypsizigus marmoreus were fractioned by hexane, chloroform, ethylacetate and buthanol. The concentrations of total polyphenolics and flavonoids in methanol extracts were investigated by colorimetric methods. The concentrations of total polyphenolics and flavonoids in methanol extract of the pileus was higher than methanol extract of the stipe. The DPPH redical scavenging activity of the pileus extract was also higher than stipe extract in methanol extract. The IC50 of DPPH redical scavenging activity of the pileus and stipe in methanol extract were 18 mg/ml and 1 10 mg/ml, respectively. The IC50 of tyrosinase inhibitory activity of the pileus and stipe in methanol extract were 500 mg/ml and 1,000 mg/ml in methanol extract. These results suggest that Hypsizigus marmoreus can be potentially used as a source of natural antioxidant agent in the cosmetic industry as well as the food, pharmaceutical and medicinal industry.

This study aims to examine the effects of taping of the ankle joint on the static and dynamic balance and gait ability of stroke patients. Twenty-six stroke patients receiving physical therapy at a hospital located in Gyeonggi-do were divided equally into a group that had taping in physical therapy and an ordinary physical therapy group. They exercised for 30 minutes each, 3 times per week for 8 weeks from June to August 2011. Romberg’s eye open and eye closed tests, limits of stability(LOS), forward and back test, timed up and go test(TUG) and 10-meter gait velocity test were performed to evaluate static balance, dynamic balance, and gait ability, respectively, prior to and 8 weeks after the intervention. Differences within each group in relation to the lapse of time were compared by a paired t-test. Differences between the two groups were compared by an independent t-test. Regarding comparison of differences within each group, all tests resulted in significant changes in both groups after the intervention (p<.05). Comparison of differences between the two groups showed that taping in the physical therapy group had significantly better test results than the ordinary physical therapy group in all measured items(p<.05). The after effects of ankle taping on stroke patients are more efficient and effective than ordinary physical therapy alone in improving balance and gait ability.

Meedon rice varieties are important for local adaptability, grain quality and market availability, and have been grown in Myanmar for centuries. Because of temporal variability and spatial heterogeneity, Meedon rice varieties in rainfed lowland areas may be diverse. However, information on diversity of Meedon rice germplasm is limited. This study was carried out to assess genetic diversity and to analyze population structure of Meedon rice germplasm conserved in Myanmar Seed Bank using SSR markers. For assessing genetic diversity, 154 accessions of Meedon rice germplasm were analyzed with nine SSR markers. A total of 86 alleles were detected with an average of 9.6 alleles per locus. All the loci were found to be polymorphic, and there were considerable genetic variation among accessions with mean values of expected heterozygosity (HE) = 0.5774 and polymorphic information content (PIC) = 0.5496. High frequency of rare alleles was identified, among which 35 unique (accession-specific) alleles were observed. Based on cluster analysis, rice accessions were mainly clustered into two groups, and as a result of model-based analysis, two distinct genetic populations and an admixture were classified. This result indicated that SSR markers have proved to be useful markers for detecting genetic diversity in Meedon rice, and the occurrence of a considerably high number of rare and unique alleles in the germplasm indicates their potentiality as a reservoir of rare genotypes for use. Unique alleles are also important because they may be diagnostic of a particular type of genotype for identification.

Adipose tissue-derived mesenchymal stem cells (ASCs) are very interesting in several laboratory animals and humans because they are easy to harvest and expand to generate millions of cells from a small quantity of fat. ASCs are known as useful materials for clinical applications in human cell therapy and as a donor cell in somatic cell nuclear transfer (SCNT). Here, we investigated if 1) minipig ASCs can be isolated, self-renewed and differentiated into multiple tissue lineages, 2) ASCs can be a suitable donor cell type for generation of cloned pig. In order to isolate ASC, adipose tissues were collected from inguinal region of a 6-year-old female minipig. The ASCs were attached to the culture dish with a fibroblast-like morphology. They expressed cell-surface marker characteristics of stem cell, underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. To investigate its potential as donor cell for cloning, we respectively carried out SCNT using ASC, adult skin fibroblast (ASF) and fetal fibroblast (FF) derived from same minipig. The ratio of blastocysts to 2-cell embryos and total cell number of blastocysts were monitored as experimental parameters. In results, cleavage and developmental competence to blastocysts rate showed no significant difference among the three groups. On the other hand, total cell numbers of blastocysts derived from ASC and FF were significantly higher than in ASF (89±7.9 and 105±5.5 vs. 57.5±5.2, respectively). Our results demonstrated that ASC have potential compared to ASF and FF in terms of the in vitro development and blastocyst formation ability. In further study, we will investigate the in vivo developmental ability of ASC as donor cell for pig cloning. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program, TS Corporation and Optifarm Solution.

Neural stem cells (NSCs) are self-renewing tripotent cell populations and have capacity of neuronal (neurons) and glial (astrocytes and oligodendrocytes) differentiation. Many researchers have reported that NSCs have therapeutic effects in neurological disease by transplantation. However, it is not easy to obtain NSCs in vitro. Recently, Yamanaka and colleagues showed that somatic cells could be reprogrammed into pluripotent state by enforcing reprogramming factors. Induced pluripotent stem (iPS) cells undergo unlimited self-renewal and have differentiation potential into various types of cells like embryonic stem cells. Direct differentiation into a specialized cell types from iPS cells hold considerable promise for regenerative medicine as well as basic research. Here, we induced differentiation of iPS cells into NSCs in vitro and in vivo, which were compared with embryonic stem (ES) cell-derived NSCs and brain derived NSCs. NSCs from ES and iPS cells were morphologically indistinguishable from brain derived NSCs and stained positive for NSCs markers Nestin and Sox2. ES cells derived NSCs were transcriptionally distinguishable from brain derived NSCs. However, global gene expression pattern were similar but distinct between iPS derived NSCs and brain derived NSCs. Moreover, iPS derived NSCs were spontaneously aggregated upon passaging, formed ES cell like colonies, and finally reactivated Oct4-GFP. The spontaneously reverted GFP-positive cells (iPS-NSC-iPS) expressed similar levels of pluripotency markers (Oct4,Nanog) to ES and iPS cells, and could form germ line chimera. One possible explanation for this phenomenon is that spontaneously re-reprogramming was associated with transgene re-activation when iPS cells were differentiated into NSCs. However, NSCs from dox-inducible iPScells could not be reprogrammed into pluripotent state without doxycycline. Taken together, iPS derived NSCs were morphologically and similar to brain derived NSCs, but differ in gene expression pattern and maintenance. * This work was supported by the Next Generation Bio-Green21 Program funded by the Rural Development Administration (Grant PJ008009).

Toxicities of 10 insecticides were examined against late third instars of Culex tritaeniorhynchus using the direct-contact mortality bioassay. Six geospatially distant field mosquitoes were collected from Chuncheon-si (designated CC-CT), Hwaseong (HS-CT), Seosan (SS-CT) Jeonju (JJ-CT), Daegu (DG-CT), and Busan (BS-CT) in the Republic of Korea (ROK). Marked regional variations of insecticide susceptibility were observed. Field populations of SS-CT, JJ-CT and DG-CT from agricultural areas showed higher to extremely higher insecticide susceptibility to pyrethroids than those of CC-CT, HS-CT and BS-CT strains from none agricultural areas. Extremely high to low levels of susceptibility were measured: bifenthrin, susceptible ratio (SR) = 2.7–896.3; β-cyfluthrin, SR = 1.8–633.3; α-cypermethrin, SR = 1.2–1,051.9; deltamethrin, SR = 1.3–711.1; permethrin, SR = 1.5–1,053.4 etofenprox, SR = 2.2–29.3; chlorfenapyr, SR = 5.1–103.6; chlorpyrifos, SR = 2.3–337.0; fenitrothion, SR = 2.0–142.3 and fenthion, SR = 1.4–186.2. Culex tritaeniorhynchus populations from rice paddies had been under heavy selection pressure due to the agricultural insecticides and that’s why the mosquito species demonstrated high resistance to pyrethroids which were used for a long time to control agricultural pests in the localities. These results indicate that careful selection and rotational use of these insecticides mayresult in continued satisfactory control against field populations of Japanese encephalitis vector mosquitoes.

This study was carried out to identify how a self-stretching exercise program affects pain for each body area, pain relief and job satisfaction for care workers. 20 of 40 care workers with musculoskeletal symptom were randomly selected and participated a self-stretching exercise program consisting of 15 motions. The intervention was done five times or more per weeks for 8 weeks and 1 session lasted within 15 minutes. 'Musculoskeletal symptom survey table' of the Korea Occupational Safety and Health Agency(KOSHA) and JDI(Job Descriptive Index) was used for pain on the musculoskeletal symptom and job satisfaction. Survey were done twice before and after the program. The result of this study showed that self-stretching exercise program group(SSPG) relieved from pain significantly in the shoulders(p<.01) and lumbar(p<.05), comparing to the non selfstretching exercise program group(NSPG). Although no significant difference on variations in the JDI appeared in SSPG, the significant reduction appeared from the colleague relationship and organization in NSPG(p<.05). SSPG showed the significant increase on variations in JDI from the job and organization comparing to NSPG. Especially, the improvement on satisfaction for the organization was shown(p<.05). Accordingly, the self-stretching exercise program for care workers can be said to positively affect the overall pain relief and increase on the JDI.

The purpose of this study was to investigate the influence of handicraft activities on hand promptness and grasp in the elderly. Subjects were comprised of 14 senior citizens between the ages of 70-85, with 7 subjects in the experiment group and 7 in the control group. Subjects in the experiment group practiced various handicrafts twice a day, while those in the control group did not participate in any special activity. The Jebsen Taylor Hand Function Test was used to evaluate the results, while a dynamometer and pinch gauge were used to measure hand promptness and grasp. The 7 senior citizens in the experiment group were able to increase their hand promptness and grasping skills. Conclusively, handicrafts can help improve hand promptness and grasp in the elderly. Furthermore, the development and improvement of such skills can have a positive influence on the daily lives of senior citizens. Such skills are expected to improve the overall neuro-function in the elderly population.

This study was to examine on the respiratory variables, heart rate and muscle activity between the static recovery and dynamic recovery after progressive resistance exercise to maximal point. Subjects were 15 students enrolled in N University. All were tested two times (static recovery and dynamic recovery) and were requested to perform a walking on a treadmill after progressive resistance exercise to maximal point. Electromyography(EMG) was used to monitor the muscle activity(TA: Tibialis Anterior, MG: Medial Gastrocnemius) during gait. CPEX-1 was used to measure the respiratory variables and heart rate. The dynamic recovery group was shown the significant lower heart rate than that of static recovery group at during gait. Respiratory rate showed statistically a significant difference. Electromyography(RMS, root mean square) showed a non-significant difference. But the dynamic recovery group of muscle activity was found highly in TA and MG. This study indicated that the dynamic recovery method evidenced more faster than the static recovery method. And this type of dynamic rest by walking can be a help of recovery after exercise.

In this case report, we investigated the effects of robot-assisted gait therapy in a chronic stroke patient using motor assessment and gait analysis. A patient who suffered from the right hemiparesis following the left corona radiata and basal ganglia infarction received 30 minutes of robot-assisted gait therapy, 3 times a week for 4 weeks. Outcome was measured using Motoricity index(MI), Fugl-Meyer assessment(FMA), modified motor assessment scale(MMAS), isometric torque, body tissue composition, 10-meter gait speed and gait analysis. After robot-assisted gait therapy, the patient showed improvement in motor functions measured by MI, FMA, MMAS, isometric torque, skeletal muscle mass, 10-meter gait speed. In gait analysis, cadence, single support time, double support time, step length, walking speed improvement in after robot-assisted gait therapy. The results of this study showed that robot-assisted gait therapy is considered to facilitate locomotor recovery of the chronic hemiparetic stroke patient.

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.

The porcine reproductive and respiratory syndrome virus (PRRSV) has three major structural proteins which designated as GP4, GP5, and M. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the GP4, GP5, and M with SUMO (Small ubiquitin-related modifier), and expressed the fused gene in Bm5 cells and silkworm larvae. Expression of the proteins were analyzed by 12% SDS-PAGE and western blotting using 6xHis tag and porcine anti-PRRSV antibodies. In results, SUMO fused proteins were expressed at a high level in Bm5 cells. The levels of protein using the silkworm larvae is higher than that using Bm5 cells. The fused protein was purified by Ni-NTA affinity chromatography. This study demonstrated that SUMO, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.