• Mass Spectrometry Letters
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• v.13 no.4
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• pp.146-151
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• 2022

Rotigotine (RTG) is a non-ergot dopamine agonist used to manage the early stage of Parkinson's disease (PD) as transdermal patch. However, the poor medication compliance of PD patients and skin issues related with repeated applications of RTG patches lead to the search for alternative formulations and it also requires appropriate analytical methods for their in vivo evaluation. Thus, here, a sensitive, efficient, and cost-effective method to determine RTG in rat plasma using liquid-liquid extraction (LLE) and multiple reaction monitoring was developed. The use of 20 µL of rat plasma for sample treatment, 8-OH-DPAT as the internal standard, and methyl tert-butyl ether as the LLE solvent in the present method gives it advantages over previous methods for the analysis of RTG in biological samples. The good analytical performance of the developed method was confirmed in specificity, linearity (the coefficient of determination ≥0.999 within 0.1-100 ng/mL), sensitivity (the lower limit of quantitation at 0.1 ng/mL), accuracy (81.00-115.05%), precision (≤10.75%), and recovery (81.00-104.48%) by following the FDA guidelines. Finally, the applicability test of the validated method to the in vivo evaluation of a RTG formulation showed that the present method is the only method which can be accurately applied to that longer than 24 hours, critical for the development of formulations with reduced dosing frequencies. Therefore, the present method could contribute to the development of new RTG formulations helpful to people suffering from PD.

• Biomolecules & Therapeutics
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• v.31 no.3
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• pp.264-275
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• 2023

Parkinson's disease (PD) is a common neurodegenerative disorder characterized by tremors, bradykinesia, and rigidity. PD is caused by loss of dopaminergic (DA) neurons in the midbrain substantia nigra (SN) and therefore, replenishment of DA neurons via stem cell-based therapy is a potential treatment option. Astrocytes are the most abundant non-neuronal cells in the central nervous system and are promising candidates for reprogramming into neuronal cells because they share a common origin with neurons. The ability of neural progenitor cells (NPCs) to proliferate and differentiate may overcome the limitations of the reduced viability and function of transplanted cells after cell replacement therapy. Achaete-scute complex homolog-like 1 (Ascl1) is a well-known neuronal-specific factor that induces various cell types such as human and mouse astrocytes and fibroblasts to differentiate into neurons. Nurr1 is involved in the differentiation and maintenance of DA neurons, and decreased Nurr1 expression is known to be a major risk factor for PD. Previous studies have shown that direct conversion of astrocytes into DA neurons and NPCs can be induced by overexpression of Ascl1 and Nurr1 and additional transcription factors genes such as superoxide dismutase 1 and SRY-box 2. Here, we demonstrate that astrocytes isolated from the ventral midbrain, the origin of SN DA neurons, can be effectively converted into DA neurons and NPCs with enhanced viability. In addition, when these NPCs are inducted to differentiate, they exhibit key characteristics of DA neurons. Thus, direct conversion of midbrain astrocytes is a possible cell therapy strategy to treat neurodegenerative diseases.

• Journal of the Korean Society of Tobacco Science
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• v.27 no.2
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• pp.212-218
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• 2005

Cigarette smoking has been known to have a few beneficial effects on some neuronal diseases such as Alzheimer's disease(AD), Parkinson's disease(PD) and prion disease by scrapie agent shows many similar properties with AD. In this respect, we investigated what biological effects are exerted by cigarette smoke exposure(CSE) in the brain of mouse infected by 87V scrapie. The scrapie agent was inoculated through stereotaxic microinjection of the homogenates of the scrapie agent infected brain into the intracerebral system in the 1M mice. The inoculation into mice typically exhibits neurochemical, physiological and histopathological characteristics of prion disease: loss of neurotransmitters and induction of astrocytosis and vacuolation in brain as well as reduction of spatial movement and loss of body weight. CSE led to alleviated the loss of body weight and also improved spatial movement of the infected mice. Most interestingly, CSE attenuated astrocytosis and vacuolation caused by scrapie infection in the brain. In addition, decreased levels of dopamine in striatal and hypothalamic regions as well as serotonin level in hippocampus caused by scrapie infection were also attenuated by exposure to cigarette smoke. These findings suggest that cigarette smoke, by its inhibition of astrocytosis and vacuolation followed by its restoration of levels of some neurotransmitters, may partly contribute to suppression in the progress of neurodegeneration caused by scrapie infection.

• Journal of Life Science
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• v.32 no.10
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• pp.771-777
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• 2022

Dopaminergic (DA) cell death in Parkinson's disease (PD) has been attributed to multiple, distinct genetic and environmental factors. In rare familial PD loss of parkin function mutations play a key role in nigral DA neuron-specific pathogenesis primarily via endoplasmic reticulum (ER) stress. In more prevalent sporadic PD, environmental exposure to pesticides has a significant epidemiological role. However, it is largely unknown how environmental exposure to xenobiotics is etiologically linked with the known etiology in familial PD. In the present study biochemical evidence for a common pathogenic mechanism between sporadic and familial PD has been identified employing the recently characterized mesencephalic DA cell line, N27-A. Dieldrin, an organochlorine pesticide epidemiologically implicated in sporadic PD, induced the markers of ER stress response such as a chaperone BiP/Grp78, heme oxygenase-1 and especially, parkin. Accordingly, dieldrin activated the ER resident Caspase-12, a mediator of ER stress-specific apoptosis, during cell death of N27-A cells. Of great interest the dieldrin-induced DA neuronal cell death was synergistically rescued by the overexpression of ER resident neuroprotective proteins, parkin and Bcl-2. The present findings implicate that accumulation of ER stress could be one of common pathogenic mechanisms in idiopathic and familial PD, and some ER proteins, such as parkin and Bcl-2 may effectively attenuate ER stress-mediated N27-A DA cell death.

• The Korean Journal of Nuclear Medicine
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• v.39 no.1
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• pp.9-14
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• 2005

Purpose: Abnormal myocardial perfusion may be caused by ventricular preexcitation, but its location, extent, severity and correlation with accessory pathway (AP) are not established. We evaluated perfusion patterns on myocardial perfusion SPECT and location of AP in patients with WPW (Wolff-Parkinson-White) syndrome. Materials and Methods: Adenosine Tc-99m MIBI or Tl-201 myocardial perfusion SPECT was performed in 11 patients with WPW syndrome. Perfusion defects (PD) were compared to AP location based on ECG with Fitzpatrick's algorithm or electrophysiologic study and radiofrequency catheter ablation. Results: Patients had atypical chest discomfort or no symptom. Risk of coronary artery disease (CAD) was below 0.1 in 11 patients using the nomogram to estimate the probability of CAD. Coronary angiography was performed in 4 patients (mid-LAD 50% in one, normal in others). In 4 patients, AP localization was done by electrophysiologic study and radiofrequency catheter ablation (RFCA). Small to large extent ($11.0{\pm}8.5%$, range:$3{\sim}35%$) and mild to moderate severity ($-71{\pm}42.7%$, range:$-2l7{\sim}-39%$) of reversible (n=9) or fixed (n=1) perfusion defects were noted. One patient with right free wall (right lateral) AP showed normal. PD locations were variable following the location of AP. One patient with left lateral wall AP was followed 6 weeks after RFCA and showed significantly decreased PD on SPECT with successful ablation. Conclusion: Myocardial perfusion defect showed variable extent, severity and location in patients with WPW syndrome. Abnormal perfusion defect showed in most of all patients, but it did not seem to be correlated specifically with location of accessory pathway and coronary artery disease. Therefore myocardial perfusion SPECT should be interpreted carefully in patients with WPW syndrome.

• The Korean Journal of Nuclear Medicine
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• v.30 no.3
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• pp.315-324
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• 1996

The purpose of this study was to compare the specific binding ratio method with model-based methods in estimating the transporter parameter $k_3/k_4$ in normal controls and Parkinson's patients with [I-123]IPT SPECT and to evaluate the usefulness of [I-123]IPT SPECT. $6.5{\pm}1.1$ mCi ($239.0{\pm}40.3$ MBq) of [$^{123}I$]IPT was intravenouly injected as a bolus into six normal controls(age:$45{\pm}13$) and seventeen patients(age:$55{\pm}8$) with Pakinson's disease(PD). The transporter parameter $k_3/k_4$ was derived using the Ichise's graphical method($R_v$) and Lassen's area ratio method($R_A$) for the dynamic IPT SPECT data without blood samples. Then, the relationships between the transporter parameter $R-v,\;R_A$ and the ratio of (BG-OCC)/OCC at 115 minutes were evaluated by linear regression analysis. $R_vs$ by Ichise's graphical method for NC and PD were $2.08{\pm}0.29$ and $0.78{\pm}0.31$, respectively. $R_As$ by Lassen's area ratio method for NC and PD were $1.48{\pm}0.16$ and $0.65{\pm}0.24$, respectively. The correlation coefficients between (BG-OCC)/OCC and $R_v$, (BG-OCC)/OCC and $R_A$, and $R_v$ and $R_A$ were 0.93, 0.90, 0.99 and their corresponding slopes were 0.54, 0.34, and 0.65, respectively. The $R_v$ and $R_A$ of NC were significantly higher than the ones of PD. That is, the $k_3/k_4$ of NC was clearly separated from the one of PD. $k_3/k_4$ showed a good correlation with the ratio of (BG-OCC)/OCC. The results indicate that the noninvasive simplified quantitative methods may be useful to measure the transporter parameter $k_3/k_4$ and the specific binding ratio method can be used for quantitative studies of dopamine transporter with [I-123]IPT SPECT in humans brains.

• Molecular & Cellular Toxicology
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• v.4 no.2
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• pp.106-112
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• 2008

Parkinson's disease (PD) progresses severely by a gradual loss of dopaminergic neurons in the substantia nigra (SN). Epidemiological studies showed that the incidences of PD were reduced by smoking of which the major component, nicotine might be neuroprotective. But the function of nicotine, which might suppress the incidences of PD, is still unknown. Fortunately, recently it was reported that a glial reaction and inflammatory processes might participate in a selective loss of dopaminergic neurons in the SN. The levels of tumour necrosis factor (TNF)-${\alpha}$ synthesised by astrocytes and microglia are elevated in striatum and cerebrospinal fluid (CSF) in PD. TNF-${\alpha}$ kills the cultured dopaminergic neurons through the apoptosis mechanism. TNF-${\alpha}$ release from glial cells may mediate progression of nigral degeneration in PD. Nicotine pretreatment considerably decreases microglial activation with significant reduction of TNF-${\alpha}$ mRNA expression and TNF-${\alpha}$ release induced by lipopholysaccharide (LPS) stimulation. Thus, this study was intended to explore the role of nicotine pretreatment to inhibit the expressions of TNF-${\alpha}$ mRNA in human fetal astrocytes (HFA) stimulated with IL-$1{\beta}$. The results are as follows: HFA were pretreated with 0.1, 1, and $10{\mu}g/mL$ of nicotine and then stimulated with IL-$1{\beta}$ (100 pg/mL) for 2h. The inhibitory effect of nicotine on expressions of TNF-${\alpha}$ mRNA in HFA with pretreated $0.1{\mu}g/mL$ of nicotine was first noted at 8hr, and the inhibitory effect was maximal at 12 h. The inhibitory effect at $1{\mu}g/mL$ of nicotine was inhibited maximal at 24 h. Cytotoxic effects of nicotine were noted above $10{\mu}g/mL$ of nicotine. Moreover, Nicotine at 0.1, 1 and $10{\mu}g/mL$concentrations significantly inhibited IL-$1{\beta}$-induced TF-${\kappa}B$ activation. Collectively, these results indicate that in activated HFA, nicotine may inhibit the expression of TNF-${\alpha}$ mRNA through the pathway which suppresses the NF-${\kappa}B$ activation. This study suggests that nicotine might be neuroprotective to dopaminergic neurons in the SN and reduce the incidences of PD.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2329-2332
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• 2007

Salsolinol, endogenous neurotoxin, is known to be involved in the pathogenesis of Parkinson's disease (PD). In the present study, we have investigated the oxidative damage of DNA induced by the reaction of salsolinol with Cu,Zn-SOD. When plasmid DNA incubated with salsolinol and Cu,Zn-SOD, DNA cleavage was proportional to the concentrations of salsolinol and Cu,Zn-SOD. The salsolinol/Cu,Zn-SOD system-mediated DNA cleavage was significantly inhibited by radical scavengers such as mannitol, ethanol and thiourea. These results indicated that free radicals might participate in DNA cleavage by the salsolinol/Cu,Zn-SOD system. Spectrophotometric study using a thiobarbituric acid showed that hydroxyl radical formation was proportional to the concentration of salsolinol and was inhibited by radical scavengers. These results indicated that hydroxyl radical generated in the reaction of salsolinol with Cu,Zn-SOD was implicated in the DNA cleavage. Catalase and copper chelators inhibited DNA cleavage and the production of hydroxyl radicals. These results suggest that DNA cleavage is mediated in the reaction of salsolinol with Cu,Zn-SOD via the generation of hydroxyl radical by a combination of the oxidation reaction of salsolinol and Fenton-like reaction of free copper ions released from oxidatively damaged SOD.

• Annals of Clinical Neurophysiology
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• v.8 no.2
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• pp.107-124
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• 2006

The basal ganglia are a group of nuclei located in the deep portion of the brain. Along with the cerebellum, the basal ganglia have a major role in controlling human voluntary movements, and their dysfunction is apparently responsible for various involuntary movements. Although the exact mechanism of how the basal ganglia control movements has yet to be clarified, the model of focused selection (through the direct pathway) and tonic inhibition (via the indirect pathway) is proposed to be a principal functional model of the basal ganglia. Parkinson's disease (PD) is classically characterized by bradykinesia, rigidity and tremor-at-rest. All features seem to be associated with dopamine depletion resulting from the degeneration of the nigrostriatal pathway, which produces reduced activity of the direct pathway and a concurrent enhancement of excitatory output from STN. This change may result in increased tonic background inhibition and reduced focused selection via the direct pathway, causing difficulties in performing voluntary movements selectively. However, it has not been possible to define a single underlying pathophysiologic mechanism that explains all parkinsonian symptoms. Here the data that give separate understanding to each of the three classic features are discussed.

• Biomedical Science Letters
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• v.17 no.2
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• pp.167-171
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• 2011

[ ${\alpha}$ ]synuclein (${\alpha}$-syn) is implicated in the pathogenesis of Parkinson's disease (PD) and other related diseases. We have previously reported that ${\alpha}$-syn binds to the cell membranes in a transient and reversible manner. However, little is known about the physiologic function and/or consequence of this association. Here, we examined whether chemically induced perturbations to the cellular membranes enhance the binding of ${\alpha}$-syn, based on hypothesis that ${\alpha}$-syn may play a role in maintenance of membrane integrity or repair. We induced membrane perturbations or alterations in ${\alpha}$-syn-overexpressing human neuroblastoma cells (SH-SY5Y) by treating the cells with hydrogen peroxide ($H_2O_2$) or oleic acid. In addition, membranes fractionated from these cells were perturbed by treating them with proteinase K or chloroform. Dynamic interaction of ${\alpha}$-syn to the membranes was analyzed by the chemical cross-linking assay that we developed in the previous study. We found that membrane interaction of ${\alpha}$-syn was increased upon treatment with membrane-perturbing reagents in a dose and time dependent manner. These results suggest that perturbations in the cellular membranes cause increased binding of ${\alpha}$-syn, and this may have significant implication in the physiological function of ${\alpha}$-syn in cells.