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.

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.

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.

Supposing that somatotype evaluation results would have significant differences between the public group with less amounts of exercises and the special group with intensive exercises for three to four times a day, this study aimed at comparing the mutual consistency between the results determined by somatotype evaluation tools such as visually calculated index(VCI), Rohrer's Index(RI) and Body Mass Index(BMI). The public, taekwondo players and judo players groups were com˗ posed of fifty persons, taekwondo players and judo players passed through VCI determination, respectively. Their height and weight were examined and analyzed with somatotype evaluation tools. Comparison of somatotype dispersion of RI and VCI showed that most women were determined by VCI as lean type but were deter˗ mined by RI as normal type. And that women were determined by VCI as fat type but were determined by RI as normal type. Therefore both men and women showed significant differences in VCI and RI. Comparison of somatotype dispersion of VCI and BMI showed that both men and women were overestimated or underestimated by VCI rather than by BMI. Comparison of somatotype dispersion of RI and BMI showed that men were less determined by BMI as lean type compared with women; both men and women less determined by BMI rather than by RI as normal type; and both men and women, in particular, were more determined by BMI as fat type but men were more determined by BMI rather than by RI as fat type. Total somatotype consistency by tools showed that VCI has the greatest possibility of determining the public group, compared with other groups as lean type and that the consistency of the three tools were relatively higher for the taekwondo players and judo players groups, compared with the public.

The purpose of this study was to determine the effects of heat appli˗ cation on the immune activities of the human body. To exam, further˗ more, the immune effect from the healthy volunteer(male:15, female:15) by monitoring changes of immune substances such as various leukocytes[total white blood cell(WBC), eosinophil, neutrophil, basophil, monocyte, and lymphocyte], a comparative study with warm water immersion(40.8±0.3℃) and infrared(250W) was carried out. The plasma analysis showed that the count of white blood cell, eosinophil, and neutrophil were elevated in warm water immersion- or infrared˗ stimulated group compared with control group. However, the count of basophil was decreased in both warm water immersion- and infrared-stimulated group than control group. Therefore, these results suggest that the thermostimulation improved immune activity.

Aujeszky’s disease (AD), also called pseudorabies, is an infectious viral disease, caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. AD affects many countries and regions in the world, causing important economic losses, mainly due to international trade restrictions. In this study, to determine the characteristics of the Aujeszky’s disease virus (ADV), NYJ strain, which was isolated from the serum of an infected pig in 1987, we investigated the nucleotide sequence and expression of the glycoproteins gB, gC, and gD using the bBpGOZA system. We found that the glycoproteins gB, gC, and gD of NYJ consisted of 2751 bp, 1443 bp, and 1203 bp, respectively. Comparison of the NYJ with the other strains revealed nucleotide sequence identity ranging from 91.tito 99.0%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The NYJ strain was formed a distinct branch with high bootstrap support. The expression of glycoprotein gD in insect cells was characterized by SDS-PAGE and Western blotting with an anti-ADV polyclonal antibody. Glycoprotein gD of approximately 45 kDa was detected. The results of this study have implications for both the taxonomy of ADV and vaccine development.

To determine the characteristics of the Korean porcine reproductive and respiratory syndrome virus (PRRSV), CA, which was isolated from the serum of an infected pig in 2006, we investigated the nucleotide sequence and expression of the structural ORFs (ORFs 2 to 7) using the bApGOZA system. We found that the structural ORFs 2 to 7 of CA consisted of 3188 nucleotides that were the same as those formed from VR-2332. Comparison of the CA with the other strains revealed nucleotide sequence identity ranging from 89.8 to 99.5%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The CA strain was closely related to the other North American genotype strains but formed a distinct branch with high bootstrap support. Additionally, expression levels of the PRRSV proteins in Sf21 cells were strong or partially weak. The results of this study have implications for both the taxonomy of PRRSV and vaccine development.

This paper describes the traditional Ondol medical care, the specific combination of ancient and contemporary examples to be articulated. As a traditional Korean culture, to be extracted from the historical perspective, it described the relationship and origins between them, From a health perspective, in accordance with the logic and pharmacologyit introduced Ondol as the important principle of health care equipment. The modern architecture of some common harmful to the body, from the perspective of Ondol, it should be improved and based on scientific proof to the theory.

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.