Background: Treadmill training is an effective intervention method for improving the walking ability of stroke patients, and taping is effective for stabilizing joints. However, taping interventions have not been implemented during treadmill training. Objectives: To examine whether treadmill training with an elastic tape or treadmill training with a non-elastic tape could be more effective in stroke patients. Design: A single blinded, randomized, controlled, comparative study. Methods: 22 stroke patients were randomly allocated to two groups: the elastic group (treadmill gait training with ankle elastic tape on the paretic side) or the non-elastic group (treadmill gait training with non-elastic tape on the paretic side). All participants performed 60 min of comprehensive rehabilitation therapy and treadmill training with an elastic tape or non-elastic tape for 20 min. Results: 10-meter walk test and timed up-and-go test results after training differed significantly from baseline in both groups (P<.05), but significantly larger gains were observed in the elastic group (10-meter walk test, -17.1%; timed up-and-go test, -18.49%; P<.05, respectively). Conclusion: Treadmill gait training with elastic tape on the affected ankle joint might be more effective at improving the walking and balancing abilities of stroke patients.

This study demonstrated a rapid and simple method for the determination of seven anions including halides and oxyhalides from the KURT underground water sample by capillary electrophoresis with UV detection. In nuclear waste disposal, some anions such as iodine, selenium, and technetium have been of great concern due to its high mobility and toxicity with a long half-life. It has been needed for a reliable analysis of anionic speciation because the high mobility of anions is easily affected by environmental conditions especially pH and salinity of underground water. Here this project is to develop a fast separation of seven anions including iodide, iodate, and selenite using capillary electrophoresis. The electroosmotic flow (EOF) was suppressed using a poly (ethyleneglycol) -coated capillary (DB-WAX capillary). As a result, anions migrated depending on their mobility under a reverse polarity condition (-15 kV) and the analysis time was within 15 min. UV detection was used at 200 nm. The RSDs for migration time were between 0.7% and 1.3% except for selenite of 5.1%. The RSDs for peak area were obtained between 2.9% and 7.4%. The calibration curves were linear from 10 to 200 mg/L with correlation coefficients greater than 0.9952. The LODs were 7.3, 10.9, 11.3, 12.9, 13.0, 13.9, and 17.4 mg/L for iodide, nitrate, bromide, selenite, bromate, tungstate and iodate. The KURT underground water sample spiked with seven anions at 50 mg/L were analyzed. The recoveries of spiked KURT sample ranged from 93.4% to 99.3%. The proposed method was successfully applied to determine seven anions in underground water sample.

Dissolution behaviors of ThO2(cr) and PuO2(cr) in synthetic groundwater were investigated at room temperature (23  2°C) under atmospheric conditions. The synthetic groundwater was prepared according to the chemical composition of the KURT-DB3 groundwater. The pH and Eh of the synthetic groundwater were pH 8.9 and 0.5 V, respectively, and the major components were Na, K, Ca, Mg, Si, Cl, SO4, F and HCO3 ions. A few mg of ThO2(cr) and PuO2(cr) powder were added in the synthetic groundwater and the concentrations of Th and Pu in supernatant were monitored for 5 months of reaction time. The concentrations of Th before and after ultracentrifugation were compared, while the solid-liquid phase separation of Pu samples could not be applied due to the small volume of sample solutions. The concentrations of Th and Pu were measured by ICP-MS and alpha spectrometry, respectively. Geochemist’s Work Bench (GWB, standard, 17.0) was applied for the modeling with ThermoChimie TDB v. 11a, which was updated with the latest NEA-TDB (vol. 14). Aqueous species distributions and solubility limiting solid phases of Th and Pu under the synthetic groundwater conditions were evaluated. The results of geochemical modeling indicate that aqueous Th-OH-CO3 ternary species and Pu(IV) species are dominant in solutions equilibrated with ThO2(s) and PuO2(am, hyd), respectively. The dissolution behaviors of ThO2(cr) and PuO2(cr) are comparable to the dissolution of ThO2(aged, logKsp = 8.5) and the oxidative dissolution of PuO2(am, hyd) in the presence of PuO2(coll, hyd), respectively.

Musculoskeletal disorders including fracture, tendonitis, osteoarthritis, and laminitis are common diseases in racehorses that can cause large economic losses in the racehorse industry. Mesenchymal stem cells (MSCs) are being applied as new clinical tools for treatment of musculoskeletal disorders of racehorses. To investigate the immunomodulatory effects of stem cell therapy, we analyzed the anti- and pro-inflammatory factors in peripheral blood mononuclear cells of racehorses before and after stem cell application using quantitative real-time RT-PCR. The expression levels of pro-inflammatory factors (CCL5, IFN-γ, IL-2, and IL-18) were decreased while those of anti-inflammatory factors (TIMP-1, IL-10, TGF-β1, and VEGF) were increased significantly after application of equine adipose tissue-derived MSCs (eAD-MSCs) to racehorses with fracture. Moreover, the expression levels of pro-inflammatory factors (IL-2, IL-18, and TNF-α) were decreased while those of anti-inflammatory factors (TIMP-1, TIMP-2, IL-10, TGF-β1, and VEGF) increased significantly after stem cell application of eAD-MSCs in racehorses with tendonitis. After evaluating immunomodulatory effects of stem cell therapy on equine musculoskeletal disorders such as fracture and tendonitis, our results showed that expression levels of pro-inflammatory factors were decreased, while those of anti-inflammatory factors increased significantly after stem cell application of eAD-MSCs. These findings suggest that the healing effects of the stem cell therapy might be due to its modulation of inflammatory factors.

Mesenchymal stem cells (MSCs), which are present in all tissues, can differentiate into cells with various specific functions. Recently, cell-based therapies using MSCs have been increasing in the veterinary research and related fields. In this study, we investigated the cellular morphology, proliferating capacities, expression of cell surface markers such as CD13, CD34, CD44, CD45, CD90, and CD105, mesodermal differentiation potentials, and expression of senescence-related markers of p53, p21, and telomerase reverse transcriptase in equine adipose tissue-derived MSCs (eAD-MSCs) after cryopreservation. The eAD-MSCs were analyzed immediately and after being frozen in liquid nitrogen for 1 year (< 1 year, G1) and more than 3 years (> 3 years, G2), respectively. After cryopreservation for 1 - 3 years, G2 eAD-MSCs showed similar cellular morphology, proliferating capacities, and expression of cell surface markers when compared with G1 eAD-MSCs. Moreover, cryopreservation did not affect the adipogenic, chondrogenic, or osteogenic differentiation potentials of G1 and G2 eAD-MSCs. Collectively, cryopreservation for (or over) 3 years maintained the stem cell phenotype and differentiation potentials of eAD-MSCs. These results will be an advantage that can be effectively used for future development of cell-based therapies.

Induced pluripotent stem cells (iPSCs) can be generated from adult cells. Somatic cells can be reprogrammed to form iPSCs by overexpressing transcription factors such as Oct4, Sox2, cMyc, and Klf4. To maintain undifferentiated state of iPSCs in vitro, cells have traditionally been maintained on mouse embryonic fibroblast feeders and passaged by enzymatic or mechanical dissociation methods. In this study, we compared the morphology and pluripotency of porcine iPSCs (piPSCs) after subsequent passaging using enzymatic and mechanical dissociation methods. Enzymatically and mechanically passaged piPSCs showed embryonic stem cell-like morphologies with compact cell adhesion and clear colony borders. In addition, alkaline phosphatase staining was positive for both enzymatically and mechanically passaged piPSCs. However, visual observation revealed that some colonies of enzymatically passaged piPSCs were spontaneously differentiated more than those of piPSCs mechanically passaged from 5 passage. Quantitative real-time RT-PCR demonstrated that enzymatically and mechanically passaged piPSCs expressed pluripotent genes such as Oct4, Sox2 and Nanog well at early passage. Immunofluorescent staining also confirmed that pluripotent markers such as Oct4, Sox2, and Nanog were positively expressed at early passage. However, expression levels of pluripotent genes in mechanically passaged piPSCs were also higher than those in enzymatically passaged piPSCs at early passage. Collectively, we found that mechanical passage method was better than enzymatic passage in terms of morphology and pluripotency of piPSCs at early passage. Further studies are needed to compare these dissociation methods with those obtained after more passages of piPSCs.

Mesenchymal stem cells (MSCs) have restricted life spans in vitro and can therefore only be expanded for a limited number of cell divisions before entering a senescent state and unequivocally stopping proliferation. Several types of cell culture systems have been used for large-scale expansion of MSCs. A recent trend in cell culture has been the change from serum-use to serum-supplement media. This study was conducted to investigate the proliferative effects of vegetable resources (VR) on equine adipose tissue-derived mesenchymal stem cells (eAD-MSCs) in the absence of serum and their possible mechanisms of action. Regulation of cell cycling is a key process involved in the fate of stem cells, including renewal and differentiation. In this study, we observed that the viability of eAD-MSCs was increased significantly when treated with VR under serum-free conditions. We also observed that expression levels of cell cycling-related proteins such as p53 and p21 were decreased, and proliferating cell nuclear antigen increased significantly in response to treatment with VR in eAD-MSCs under serum-free conditions. Furthermore, expression levels of cell survival-related proteins were increased in response to treatment with VR in eAD-MSCs under serum-free conditions. Therefore, our results suggest that VR promotes proliferation of eAD-MSCs under serum-free conditions.

Mesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several cell lineages, which has implications for cell therapy and reproductive biotechnologies. Although MSCs have been isolated from many species, including humans and animals, there is limited data on MSCs from large ruminants, such as bovines. In this study, we tried to isolate and characterize bovine tongue tissue-derived MSCs (boT-MSCs) by investigating phenotype morphology, performing proliferation properties, and determining cell surface marker expression patterns, self-renewal, and differentiation potentials. As a result, the boT-MSCs were successfully isolated by collagenase digestion and maintained proliferative capacity until 20 passages. Moreover, the boT-MSCs expressed pluripotency markers (OCT3/4, SOX2, and NANOG) and MSC-specific surface markers including CD44, CD90, and CD105, but not CD45 and MHC-II. The boT-MSCs could also differentiate into mesodermal (adipocyte, osteocyte, and chondrocyte) cell lineages. Our results suggest that the tongues of bovines could be used as a source of MSCs.

Mesenchymal stem cells (MSCs) are primary candidates for cell therapy and tissue engineering applications. A two-dimensional (2D) culture system is typically used for cell growth, but that method affects the characteristics of stem cells. The physiological cell environment connects cells not only to each other, but also to the extracellular matrix providing mechanical support, exposing the entire cell surface, and opening signaling pathways. The hanging drop method is the most widely used 3D culture method for spheroid formation. In this study, we investigated the relationship between spheroid size and changes in gene expression to determine the optimum spheroid size for use in tissue engineering. The expression levels of stemness factors such as NANOG, OCT4, and SOX2, angiogenic factors such as VEGF and IL-8, and osteogenic factors such as COX2 and TGF-β1 increased with spheroid size in the respective spheroid formation groups unlike the responses in their monolayer groups. Therefore, our results indicate that spheroid formation through the hanging drop method can increase the efficiency of MSCs-based tissue engineering over that obtained via traditional 2D cell culture systems.

Mesenchymal stem cells (MSCs) have been researched for use in biomedical applications, particularly for cell-based therapies and regenerative medicine due to their self-renewing capacity and ability to differentiate into multiple cell types such as adipose, bone, and tendon tissues. Cryopreservation of MSCs is a common preservation method that is advantageous for cellular therapies in human and veterinary medicine. Adipose tissue-derived cells have been shown to maintain their properties after cryopreservation. In this study, we investigated the morphology, proliferation (cumulative population doubling level and doubling time), cell surface markers (CD34, CD90, and CD105), and ability to differentiate into adipose, bone, and cartilage tissues in vitro of equine adipose tissue-derived MSCs (eAD-MSCs) and miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with and without long-term cryopreservation. The eAD-MSCs and mpAD-MSCs were analyzed immediately and after being frozen in liquid nitrogen for 3 years and 2 years, respectively. Cryopreserved eAD-MSCs maintained their morphology, proliferation rate, and cell surface markers compared with fresh cells. With the exception of proliferation rate, cryopreserved mpAD-MSCs also maintained their fresh cell characteristics. The proliferation rate of cryopreserved mpAD-MSCs was higher than that for fresh cells. Cryopreservation did not change the adipogenic, chondrogenic, or osteogenic differentiation potentials of eAD-MSCs and mpAD-MSCs. In summary, long-term cryopreservation maintains the cell phenotype and differentiation ability of eAD-MSCs and mpAD-MSCs. These results might be useful when developing veterinary medicine and clinical applications.

Mesenchymal stem cells (MSCs) are multipotent stem cells, which can be induced to differentiate into several cells. MSCs are also reported to possess immunomodulatory properties through secretion of inflammatory cytokines and generation of regulatory T (Treg) cells. Treg cells play an important role in allergic disorders, including atopic dermatitis. We examined the immunomodulatory effects of canine adipose tissue derived-MSCs (cAD-MSCs) in 3 groups: Group 1, untreated normal dog; Group 2, dogs with Dermatophagoides farinae ointment-induced atopic dermatitis; and Group 3, dogs with atopic dermatitis. Canine peripheral blood mononuclear cells (PBMCs) were collected from each group and co-cultured with cAD-MSCs. After co-culturing, PBMCs were separated and the expression of Treg cells was examined by flow cytometry. According to the results, the populations of Treg cells were increased in 3 ex vivo experimental groups, co-cultured with cAD-MSCs. These results would be important for the application of MSCs in clinical trials.

Mesenchymal stem cells (MSCs) are an attractive source for cell therapy, as they have the potential for differentiation into multi-lineage cells. Adipose tissue is a safe source due to its easy extraction and abundant resource, with minimal risk to the organ donor. In this study, we attempted to correlate the harvest yield and resulting multipotency of feline adipose tissue-derived mesenchymal stem cells (fAD-MSCs) in accordance with processing time. fAD-MSCs were individually isolated from the abdominal adipose tissues of 6 felines. They were divided into two groups, based on their processing times – Group 1: 0~1 day after adipose tissue harvesting; Group 2: more than 3 days after adipose tissue harvesting. In both groups, the proliferation capacity was analyzed using the cumulative population doubling level (CPDL) calculation assay. The expression levels of MSC-specific markers and differentiation potentials into mesodermal cell lineages were also evaluated. We observed that fAD-MSC isolation yields and CPDL were excellent in Group 1 compared with Group 2. We also found that the differentiation potential-specific genes (ACAN and OPN) were strongly expressed in Group 1 compared with Group 2. These results suggest that for the clinical treatments of feline diseases, fAD-MSCs should be isolated within 1 day after adipose tissue harvesting.