Young-Hwan Ahn

Autologous Mesenchymal Stem Cell Therapy Delays the Progression of Neurological Deficits in Patients With Multiple System Atrophy

We evaluated the feasibility and safety of therapy with mesenchymal stem cells (MSCs) through consecutively intra‐arterial and three repeated intravenous injections and compared the long‐term prognosis between MSC‐treated (n=11) and control multiple system atrophy (MSA) patients (n=18). The MSC‐treated patients showed significantly greater improvement on the unified MSA rating scale (UMSARS) than the control patients at all visits throughout the 12‐month study period. Orthostasis in UMSARS I items and cerebellar dysfunction‐related items of UMSARS II items were significantly different in favor of MSC treatment compared to controls. Serial positron emission tomography scan in the MSC‐treated group showed that increased fluorodeoxyglucose uptake from baseline was noted in cerebellum and frontal white matters. No serious adverse effects related to MSC therapy occurred. This study demonstrated that MSC therapy in patients with MSA was safe and delayed the progression of neurological deficits with achievement of functional improvement in the follow‐up period.

Brain Transplantation of Neural Stem Cells Cotransduced with Tyrosine Hydroxylase and GTP Cyclohydrolase 1 in Parkinsonian Rats

Neural stem cells (NSCs) of the central nervous system (CNS) recently have attracted a great deal of interest not only because of their importance in basic research on neural development, but also in terms of their therapeutic potential in neurological diseases, such as Parkinsons disease (PD). To examine if genetically modified NSCs are a suitable source for the cell and gene therapy of PD, an immortalized mouse NSC line, C17.2, was transduced with tyrosine hydroxylase (TH) gene and with GTP cyclohydrolase 1 (GTPCH1) gene, which are important enzymes in dopamine biosynthesis. The expression of TH in transduced C17.2-THGC cells was confirmed by RT-PCR, Western blot analysis, and immunocytochemistry, and expression of GTPCH1 by RT-PCR. The level of L-DOPA released by C17.2-THGC cells, as determined by HPLC assay, was 3793 pmol/106 cells, which is 760-fold higher than that produced by C17.2-TH cells, indicating that GTPCH1 expression is important for L-DOPA production by transduced C17.2 cells. Following the implantation of C17.2-THGcC NSCs into the striata of parkinsonian rats, a marked improvement in amphetamine-induced turning behavior was observed in parkinsonian rats grafted with C17.2-THGC cells but not in the control rats grafted with C17.2 cells. These results indicate that genetically modified NSCs grafted into the brain of the parkinsonian rats are capable of survival, migration, and neuronal differentiation. Collectively, these results suggest that NSCs have great potential as a source of cells for cell therapy and an effective vehicle for therapeutic gene transfer in Parkinsons disease.

Simultaneous clinical monitoring of lactic acid, pyruvic acid and ketone bodies in plasma as methoxime/tert-butyldimethylsilyl derivatives by gas chromatography–mass spectrometry in selected ion monitoring mode

Simultaneous determination of lactic acid, pyruvic acid, 3-hydroxybutyric acid and acetoacetic acid for clinical monitoring of lactic acidosis and ketone body formation in human plasma (20 microL) was performed by gas chromatography-mass spectrometry in selected ion monitoring (SIM) mode after generating methoxime/tert-butyldimethylsilyl derivatives. All of the targeted carboxylic acids were detected by characteristic fragment ions, which permitted sensitive and selective identification in the presence of co-extracted free fatty acids and other acidic metabolites at much higher levels. The method was linear (r>or=0.9991), reproducible (% relative standard deviation=1.2-5.8), and accurate (% relative error=-7.2-7.6), with detection limits of 0.05-1.7 ng/mL. This rapid, accurate and selective method using minimal plasma samples (20 microL) is useful in the clinical monitoring of lactic acidosis and ketone body formation in plasma.

Increased fiber outgrowth from xeno-transplanted human embryonic dopaminergic neurons with co-implants of polymer-encapsulated genetically modified cells releasing glial cell line-derived neurotrophic factor.

We investigated whether a continuous supply of glial cell line-derived neurotrophic factor (GDNF) via encapsulated genetically modified cells can promote survival and fiber outgrowth from xenotransplanted human dopaminergic neurons. Cells genetically engineered to continuously secrete GDNF were confined in hollow fiber-based macrocapsules. Each hemiparkinsonian rat received either a single C2C12-hGDNF capsule (n=8) or a C2C12-control capsule (n=8) concomitantly with human embryonic ventral mesencephalic cell suspension transplants. Our results show that fiber outgrowth in the area between the capsule and the graft is more extensive in rats with GDNF-releasing capsules than in rats with control capsules. We suggest that continuous and safe delivery of GDNF to the brain could be a potential way to optimize neural transplantation as a therapy for Parkinsons disease.

Polyamine patterns in the cerebrospinal fluid of patients with Parkinson's disease and multiple system atrophy.

BACKGROUND Polyamines (PAs) are important modulators of physiological condition, and are associated with neurodegenerative disease. Thus, we investigated the change of PA concentration in the cerebrospinal fluid (CSF) of patients with Parkinsons disease (PD) and multiple system atrophy (MSA). METHODS CSF samples from patients with PD and MSA were examined by gas chromatography-mass spectrometry in selected ion monitoring mode using N-ethoxycarbonyl/N-pentafluoropropyonyl derivatives. RESULTS PA concentrations were significantly different in patients with PD and MSA compared with those in the normal group. In the PD group, as compared with the MSA group, concentrations of putrescine, N(1)-acetylspermidine, and putrescine spermidine(-)(1) were significantly increased, whereas the concentration of spermidine was significantly reduced. CONCLUSIONS These results could be helpful for understanding the complexity of biochemical events in patients with PD and MSA, and may serve as metabolic markers for diagnosis of PD and MSA.

Sensitive detection of yeast using terahertz slot antennas

We demonstrated sensitive detection of individual yeast cells and yeast films by using slot antenna arrays operating in the terahertz frequency range. Microorganisms located at the slot area cause a shift in the resonant frequency of the THz transmission. The shift was investigated as a function of the surface number density for a set of devices fabricated on different substrates. In particular, sensors fabricated on a substrate with relatively low permittivity demonstrate higher sensitivity. The frequency shift decreases with increasing slot antenna width for a fixed coverage of yeast film, indicating a field enhancement effect. Furthermore, the vertical range of the effective sensing volume has been studied by varying the thickness of the yeast film. The resonant frequency shift saturates at 3.5 μm for a slot width of 2 μm. In addition, the results of finite-difference time-domain simulations are in good agreement with our experimental data.

Gas chromatographic-mass spectrometric analyses of cholesterol and its precursors in rat plasma as tert-butyldimethylsilyl derivatives

BACKGROUND Cholesterol and its metabolic precursors occurring in metabolic pathways are important biochemical indicators in pathological conditions. METHODS A method for the simultaneous determination of cholesterol and its metabolic precursors, such as lanosterol and 7-dehydrocholesterol, in rat plasma is demonstrated. It involves their extraction after saponification, followed by conversion to tert-butyldimethylsilyl (TBDMS) derivatives for analysis by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode (GC-SIM-MS). RESULTS The characteristic fragment ions of [M-57], m/z 443, 483, and 441 permitted the accurate and selective detection of cholesterol and its precursors in rat plasma. The whole procedure of TBDMS derivatization, with subsequent GC-SIM-MS analysis, was linear (r>or=0.9994), reproducible (% relative standard deviation=2.2 to 7.5), and accurate (% relative error=-5.6 to 7.7), with detection limits of 0.02 to 0.07 ng/ml. Recoveries were measured to be ranged from 89.5 to 95.4%. CONCLUSION The present method was useful for the quantification of cholesterol and its precursors in rat plasma samples of 1 microl.

Elevated homocysteine by levodopa is detrimental to neurogenesis in parkinsonian model.

Background Modulation of neurogenesis that acts as an endogenous repair mechanism would have a significant impact on future therapeutic strategies for Parkinson’s disease (PD). Several studies demonstrated dopaminergic modulation of neurogenesis in the subventricular zone (SVZ) of the adult brain. Levodopa, the gold standard therapy for PD, causes an increase in homocysteine levels that induces neuronal death via N-methyl-D-aspartate (NMDA) receptor. The present study investigated whether elevated homocysteine by levodopa treatment in a parkinsonian model would modulate neurogenesis via NMDA receptor signal cascade and compared the effect of levodopa and pramipexol (PPX) on neurogenic activity. Methodology/Principal Findings Neurogenesis was assessed in vitro using neural progenitor cells (NPCs) isolated from the SVZ and in vivo with the BrdU-injected animal model of PD using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Modulation of homocysteine levels was evaluated using co-cultures of NPCs and astrocytes and PD animals. Immunochemical and Western blot analyses were used to measure neurogenesis and determine the cell death signaling. Levodopa treatment increased release of homocysteine on astrocytes culture media as well as in plasma and brain of PD animals. Increased homocysteine by levodopa led to increased apoptosis of NPCs through the NMDA receptor-dependent the extracellular signal-regulated kinase (ERK) signaling pathways. The administration of a NMDA antagonist significantly attenuated apoptotic cell death in levodopa-treated NPCs and markedly increased the number of BrdU-positive cells in the SVZ of levodopa-treated PD animals. Comparative analysis revealed that PPX treatment significantly increased the number of NPCs and BrdU-positive cells in the SVZ of PD animals compared to levodopa treatment. Our present study demonstrated that increased homocysteine by levodopa has a detrimental effect on neurogenesis through NMDA receptor-mediated ERK signaling pathway. Conclusions/Significance Modulation of levodopa-induced elevated homocysteine by NMDA antagonist or dopamine agonist has a clinical relevance for PD treatment in terms of adult neurogenesis.

The Potential Utility of Iodinated Contrast Media (ICM) Skin Testing in Patients with ICM Hypersensitivity

Both immediate and delayed hypersensitivity reactions to iodinated contrast media (ICM) are relatively common. However, there are few data to determine the clinical utility of immunologic evaluation of ICM. To evaluate the utility of ICM skin testing in patients with ICM hypersensitivity, 23 patients (17 immediate and 6 delayed reactions) were enrolled from 3 university hospitals in Korea. With 6 commonly used ICM including iopromide, iohexol, ioversol, iomeprol, iopamidol and iodixanol, skin prick (SPT), intradermal (IDT) and patch tests were performed. Of 10 patients with anaphylaxis, 3 (30.0%) and 6 (60.0%) were positive respectively on SPTs and IDTs with the culprit ICM. Three of 6 patients with urticaria showed positive IDTs. In total, 11 (64.7%) had positive on either SPT or IDT. Three of 6 patients with delayed rashes had positive response to patch test and/or delayed IDT. Among 5 patients (3 anaphylaxis, 1 urticaria and 1 delayed rash) taken subsequent radiological examinations, 3 patients administered safe alternatives according to the results of skin testing had no adverse reaction. However, anaphylaxis developed in the other 2 patients administered the culprit ICM again. With 64.7% (11/17) and 50% (3/6) of the sensitivities of corresponding allergic skin tests with culprit ICM for immediate and delayed hypersensitivity reactions, the present study suggests that skin tests is useful for the diagnosis of ICM hypersensitivity and for selecting safe ICM and preventing a recurrence of anaphylaxis caused by the same ICM. Graphical Abstract

Sensing viruses using terahertz nano-gap metamaterials

We demonstrate highly sensitive detection of viruses using terahertz split-ring resonators with various capacitive gap widths. Two types of viruses, with sizes ranging from 60 nm (PRD1) to 30 nm (MS2), were detected at low densities on the metamaterial surface. The dielectric constants of the virus layers in the THz frequency range were first measured using thick films, and the large values found identified them as efficient target substances for dielectric sensing. We observed the resonance-frequency shift of the THz metamaterial following deposition of the viruses on the surface at low-density. The resonance shift was higher for the MS2 virus, which has a relatively large dielectric constant. The frequency shift increases with surface density until saturation and the sensitivity is then obtained from the initial slope. Significantly, the sensitivity increases by about 13 times as the gap width in the metamaterials is decreased from 3 µm to 200 nm. This results from a combination of size-related factors, leading to field enhancement accompanying strong field localization.