Jiyeon Lim

Association of DRD3 and GRIN2B with impulse control and related behaviors in Parkinson's disease†

We aimed to assess whether allelic variants of dopamine receptor, glutamate receptor, and serotonin transporter genes are associated with the appearance of impulse control and related behaviors (ICRB) in Parkinsons disease (PD) with dopamine replacement therapy (DRT). We surveyed ICRB in consecutive Korean patients with PD who were treated with stable DRT using modified Minnesota Impulsive Disorders Interview over a period of 4 months. In the 404 patients who completed the interview and the 559 Korean healthy normal controls, genotyping was performed for variants of the DRD3 p.S9G, DRD2 Taq1A, GRIN2B c.366C>G, c.2664C>T and c.‐200T>G, and the promoter region of the serotonin transporter gene (5‐HTTLPR). Behavioral abnormalities suggestive of ICRB including compulsive buying, gambling, sexual behavior and eating, and punding, were present in 14.4% of the patients. Variants of DRD2 and 5‐HTTLPR were not associated with the risk of developing ICRB. However, the AA genotype of DRD3 p.S9G and the CC genotype of GRIN2B c.366C>G were more frequent in patients with ICRB than in nonaffected patients (odds ratio [OR] = 2.21, P = 0.0094; and 2.14, P = 0.0087, after adjusting for age and sex). After controlling for clinical variables in the multivariate analysis, carriage of either AA genotype of DRD3 or CC genotype of GRIN2B was identified as an independent risk factor for ICRB (adjusted OR: 2.57, P = 0.0087). Variants of DRD3 p.S9G and GRIN2B c.366C>G may be associated with the appearance of ICRB in PD.

Let-7 microRNA inhibits the proliferation of human glioblastoma cells

MicroRNAs (miRNAs) are small noncoding RNAs comprising 21–23 nucleotides that regulate gene expression by transcriptionally repressing their complementary mRNAs. In particular, let-7 miRNA has been postulated to function as a tumor suppressor in various cancer cells, but not yet in glioblastoma. In this study, we investigated the anti-tumorigenic effect of let-7 miRNA in glioblastoma cells. Human glioblastoma cells (U251 or U87 cells) were transfected with let-7 miRNA and assayed for in-vitro proliferation, migration, and in-vivo tumor formation. Transfection of let-7 miRNA reduced expression of pan-RAS, N-RAS, and K-RAS in the glioblastoma cells. Let-7 miRNA also reduced the in-vitro proliferation and migration of the cells, and reduced the sizes of the tumors produced after transplantation into nude mice. However, let-7 miRNA exerted no effect on the proliferation of normal human astrocytes. These results indicate that let-7 miRNA has an anti-tumorigenic effect on glioblastoma cells, and suggest possible use of let-7 miRNA for treating glioblastoma.

Electroacupuncture enhances motor recovery performance with brain-derived neurotrophic factor expression in rats with cerebral infarction

Objective Electroacupuncture (EA) is a traditional medicine in patients with post-stroke rehabilitation. Brain-derived neurotrophic factor (BDNF) is a potent growth factor involved in recovery following cerebral injury. The aim of the present study was to investigate whether EA increases BDNF levels and facilitates functional recovery. Methods Occlusion of the middle cerebral artery was performed in rats (N=12) followed by reperfusion. EA was applied at the GV20 (Baihui) acupoint. Motor and sensory functions were monitored on the Garcia scale for 2 weeks. Expressions of BDNF and receptor tyrosine kinase B (trkB) were determined by immunoblotting and immunohistochemistry. Results Improvement of Garcia scores, particularly in motor performance, were noted in the group with EA stimulation (p<0.05). With EA application, BDNF was elevated in the ischaemic hemisphere with increased numbers of BDNF(+) cells. Increased expression of trkB was also detected. Conclusion These results indicate that EA at GV20 improves motor recovery and stimulates BDNF/trkB expression in rats with cerebral ischaemia.

Extracts of Adipose Derived Stem Cells Slows Progression in the R6/2 Model of Huntington's Disease

Stem cell therapy is a promising treatment for incurable disorders including Huntingtons disease (HD). Adipose-derived stem cell (ASC) is an easily available source of stem cells. Since ASCs can be differentiated into nervous stem cells, it has clinically feasible potential for neurodegenerative disease. In addition, ASCs secrete various anti-apoptotic growth factors, which improve the symptoms of disease from transplanted ASCs. Thus, cell-free extracts of ASCs (ASCs-E) could be a potential candidate for treatment of HD. Here, we investigated effects of ASCs-E on R6/2 HD mouse model and neuronal cells. In R6/2 HD model, injection of ASCs-E improved the performance in Rotarod test. ASCs-E also ameliorated striatal atrophy and mutant huntingtin aggregation in the striatum. In Western blot increased expressions of p-Akt, p-CREB and PGC1α were noted by injection of ASCs-E, when comparing to the R6/2 HD model. Neuro2A neuroblastoma cells treated with ASCs-E showed increased expression of p-CREB and PGC1α. In conclusion, ASCs-E delayed disease progression in animal model of HD by restoring of CREB-PGC1α pathway and could be a potential resource for treatment of HD.

Sun Ginseng Protects Endothelial Progenitor Cells From Senescence Associated Apoptosis

Endothelial progenitor cells (EPC) are a population of cells that circulate in the blood stream. They play a role in angiogenesis and, therefore, can be prognostic markers of vascular repair. Ginsenoside Rg3 prevents endothelial cell apoptosis through the inhibition of the mitochondrial caspase pathway. It also affects estrogen activity, which reduces EPC senescence. Sun ginseng (SG), which is heat-processed ginseng, has a high content of ginsenosides. The purpose of this study was to investigate the protective effects of SG on senescence-associated apoptosis in EPCs. In order to isolate EPCs, mononuclear cells of human blood buffy coats were cultured and characterized by their uptake of acetylated low-density lipoprotein (acLDL) and their binding of Ulex europaeus agglutinin I (ulex-lectin). Flow cytometry with annexin-V staining was performed in order to assess early and late apoptosis. Senescence was determined by β-galactosidase (β-gal) staining. Staining with 4′-6-Diamidino-2-phenylindole verified that most adherent cells (93±2.7%) were acLDL-positive and ulex-lectin-positive. The percentage of β-gal-positive EPCs was decreased from 93.8±2.0% to 62.5±3.6% by SG treatment. A fluorescence-activated cell sorter (FACS) analysis showed that 4.9% of EPCs were late apoptotic in controls. Sun ginseng decreased the apoptotic cell population by 39% in the late stage of apoptosis from control baseline levels. In conclusion, these results show antisenescent and antiapoptotic effects of SG in human-derived EPCs, indicating that SG can enhance EPC-mediated repair mechanisms.

Transplantation of patient-derived adipose stem cells in YAC128 Huntington's disease transgenic mice.

Huntington’s disease (HD) is a genetic neurodegenerative disorder caused by abnormal expansion of CAG in the huntingtin gene. In R6/2 HD transgenic mice, human adipose-derived stem cells (ASCs) can slow disease progression via secretion of multiple paracrine growth factors. In order to prompt autologous ASCs transplantation in HD, we isolated ASCs from subcutaneous adipose tissues from a HD patient and a normal volunteer. ASCs were grown in two different types of stem cell culture media, EGM-2MV (endothelial growth medium-2 MV) or mesenchymal culture medium (MesenPRO). Cell-surface markers CD13, CD29, CD31, CD34, and CD44 were characterized by flow cytometry. BDNF, HGF, IGF, LIF, NGF, and VEGF expressions were determined by RT-PCR. Cell surface markers for HD ASCs were similar to those for normal ASCs. HD ASCs expressed multiple growth factors, and were similar to normal ASCs, except for NGF; however, they can be altered by culture medium. ASCs were transplanted in bilateral striata of 8 month-old YAC128 mice. At 12 months of age, normal ASCs reduced striatal atrophy, while HD ASCs failed to prevent it. Compared to the control YAC128 group, no improvement in Rotarod performance was observed in any of the transplanted YAC128 mice. However, when normal ASCs were transplanted at 12 months, Rotarod performance was maintained for 4 weeks, with detectable transplanted cells in the striatum and periventricular area. In summary, cultured HD patient-derived ASCs express multiple growth factors with the same cell surface markers as those of normal ASCs in vitro. The efficacy of ASCs transplantation in YAC128 transgenic models can be modified, depending on the time window.

Altered Expression of miR-202 in Cerebellum of Multiple-System Atrophy

Cerebellar degeneration is a devastating manifestation of cerebellar-type multiple-system atrophy (MSA), a rapidly progressive neurodegenerative disease, and the exact pathogenesis is unknown. Here, we examined the expression of micro-RNAs (miRNAs), which are short noncoding RNAs, in the cerebellum of MSA and the key target genes. miRNA microarray found 11 miRNAs with significantly different expression in MSA cerebellum compared to cerebellum from age-, sex-, and postmortem interval-matched controls. miR-202 was the most upregulated in the MSA samples. In silico analysis, followed by target gene luciferase assay, in vitro transfection, and Western blotting in human samples showed that miR-202 downregulates Oct1 (Pou2f1), a transcription factor expressed in cerebellar Purkinje cells. Transfection of Neuro-2a cells with miR-202 enhanced oxidative stress-induced cell death, and an antagomir to miR-202 inhibited this effect of miR-202. This study provides novel insight into the role of miRNA in cerebellar degeneration and suggests that miR-202 is a key miRNA mediating the pathogenesis of MSA.

Adipose-derived stem cells extract has a proliferative effect on myogenic progenitors

Finding an effective method to regenerate muscle is a growing issue in the orthopedic field. Platelet-rich plasma (PRP) has recently been considered for therapeutic use due to its capacity to induce proliferation of myogenic progenitor cells (MPCs). Adipose-derived stem cells (ASCs) and its extract are regarded as a promising treatment for various disorders within the orthopedic field but their therapeutic relevance in the muscle regeneration is poorly investigated. In this study, rabbit MPCs were cultured from the supraspinatus of rabbit and characterized by myogenic markers. To investigate the paracrine effect of ASCs on MPCs, coculture experiments were performed. In order to see the anabolic effect of ASC-extracts (ASC-ex) in MPCs, cell proliferation assays were performed and compared with the PRP-added condition. Coculture experiment showed ASCs had an anabolic paracrine effect on proliferation of MPCs. PRP had a positive effect on proliferation of MPCs when compared to the control (100 ± 7.4% vs 195.2 ± 19.2%, p < 0.001); however, ASC-ex promoted greater proliferation than the PRP condition (467.3 ± 38.7%, p < 0.001 compared with PRP). Similarly, in C2C12 cells, PRP showed an increased rate when compared to the control (100 ± 5.9% vs 205.1 ± 45.4%, p < 0.001), and treatment of ASC-ex showed dramatic increase in proliferation (335.9 ± 37.8%, p < 0.001 compared with PRP). ASC-ex had positive effect on expanding MPCs of rabbit and myoblast cell line, and its capacity to induce proliferation was notably stronger than that of PRP. In conclusion, the study suggests that rabbit ASC-ex have stronger proliferative effect on MPCs than rabbit PRP. Thus, ASC-ex could be a therapeutic candidate for muscle regeneration by activation of endogenous MPCs.

Single-Dose Gadoterate Meglumine for 3T Late Gadolinium Enhancement MRI for the Assessment of Chronic Myocardial Infarction: Intra-Individual Comparison with Conventional Double-Dose 1.5T MRI

Objective To intra-individually compare 3T magnetic resonance (MR) images obtained with one dose gadoterate meglumine to 1.5T MR using conventional double dose for assessment of chronic myocardial infarction. Materials and Methods Sixteen patients diagnosed with chronic myocardial infarctions were examined on single-dose 3T MR within two weeks after undergoing double-dose 1.5T MR. Representative short-axis images were acquired at three points after administration of gadoterate meglumine. Contrast-to-noise ratios between infarcted and normal myocardium (CNRinfarct-normal) and between infarct and left ventricular cavity (CNRinfarct-LVC) were calculated and compared intra-individually at each temporal scan. Additionally, two independent readers assessed relative infarct size semi-automatically and inter-observer reproducibility was evaluated using intraclass correlation coefficient. Results While higher CNRinfarct-normal was revealed at single-dose 3T at only 10 minutes scan (p = 0.047), the CNRinfarct-LVC was higher at single-dose 3T MR at each temporal scan (all, p < 0.05). Measurement of relative infarct size was not significantly different between both examinations for both observers (all, p > 0.05). However, inter-observer reproducibility was higher at single-dose 3T MR (all, p < 0.05). Conclusion Single-dose 3T MR is as effective as double-dose 1.5T MR for delineation of infarcted myocardium while being superior in detection of infarcted myocardium from the blood cavity, and provides better reproducibility for infarct size quantification.

Incidentally detected atherosclerosis in the abdominal aorta or its major branches on computed tomography is highly associated with coronary heart disease in asymptomatic adults

BACKGROUND Atherosclerotic lesions in the abdominal aorta or its major branches are often incidentally detected on abdominal CT. However, clinical implications and optimal subsequent management are mostly left undetermined. METHODS Consecutive, asymptomatic adults (age≥30) who underwent both abdominal CT and coronary computed tomography angiography as part of a self-referred health check-up were investigated (n = 1494). RESULTS Adjusted for cardiovascular risk factors, abdominal atherosclerotic lesions with stenosis<25% were associated with significant coronary stenosis, especially in the abdominal aorta (adjusted odds ratio [aOR] 3.37, 95% confidence interval [CI] 0.99-11.45) and any common iliac artery (aOR 2.99, 95% CI 1.43-6.26). The association was higher in atherosclerotic lesions with stenosis≥25%, respectively (aOR 16.39, 95% CI 4.00-67.11; aOR 7.32, 95% CI 2.84-18.86). Furthermore, any major abdominal artery stenosis added predictive value to significant coronary stenosis (area under the receiver operating curve: 0.7598 vs. 0.8019, P < 0.001). The extent of arterial territory involvement was associated with the presence of significant coronary stenoses (P for trend <0.001). CONCLUSION Stenotic atherosclerotic lesions in the abdominal aorta or its major branches incidentally detected on abdominal CT are relatively prevalent and carry high risk for asymptomatic coronary arterial disease.