Limamanen Phom has completed M.Sc. Zoology from Nagaland University (Central), Nagaland, India. He worked as Research Fellow in the Department of Biotechnology (Government of India) funded research project entitled, Neuroprotective Effects of Natural Products with Antiinflammatory and Antioxidative Properties in Drosophila Model of Parkinson’s Disease and has shown that Curcumin’s neuroprotective efficacy in Drosophila model of Idiopathic Parkinson’s Disease is phase specific which has therapeutic implications for human. Presently, he is pursuing Ph.D under the supervision of Dr. Sarat Chandra Yenisetti.

Selective degeneration of dopaminergic neurons in the substantia nigra underlies the basic motor impairments of Parkinson’s disease (PD). Curcumin has been used for centuries in traditional medicines in India. Our aim is to understand the efficacy of genotropic drug curcumin as a neuroprotective agent in PD. Analysis of different developmental stages in model organisms revealed that they are characterized by different patterns of gene expression which is similar to that of developmental stages of human. Genotropic drugs would be effective only during those life cycle stages for which their target molecules are available. Hence there exists a possibility that targets of genotropic compounds such as curcumin may not be present in all life stages. However, no reports are available in PD models illustrating the efficacy of curcumin in later phases of adult life. This is important because this is the period during which late-onset disorders such as idiopathic PD set in. To understand this paradigm, we tested the protective efficacy of curcumin in different growth stages (early, late health stage, and transition phase) in adult Drosophila flies. Results showed that it can rescue the motor defects during early stages of life but is ineffective at later phases. This observation was substantiated with the finding that curcumin treatment could replenish depleted brain dopamine levels in the PD model only during early stages of life cycle, clearly suggesting its limitation as a therapeutic agent in late-onset neurodegenerative disorders such as PD.

Huamin Xu, Qingdao University, China

Rosmarinic acid (RA) is a naturally occurring polyphenolic compound, and is composed of caffeic acid and danshensu. Our previous studies have confirmed RA could protect against 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenyl pyridine cation (MPP+) induced cell injury. Improving evidence showed iron-induced α-synuclein aggregation played important roles in the etiology of Parkinson's disease (PD). However, whether RA could protect dopaminergic neurons through inhibiting the aggregation of α-synuclein in PD is unclear. And the regulation mechanisms underlying this inhibition were not elucidated. Therefore, the experiment proposed to explore the effects of rosmarinic acid against iron-induced α-synuclein aggregation in dopaminergic cells and elucidate the possible mechanisms in the SK-N-SH cells. Results showed iron could reduce the mitochondrial transmembrane potential (△Ψm) and induce α-synuclein aggregation in the SK-N-SH cells. In accordance with iron responsive element/iron regulatory protein (IRE/IRP) system, iron could increase the mRNA levels of α-synuclein. And results also showed that RA pretreatment could restore the ΔΨm reduction induced by iron and alleviate iron induced α-synuclein aggregation. Further results showed RA pretreatment could inhibit iron induced α-synuclein aggregation by up-regulating hemeoxygenase-1 (HO-1). In addition, RA pretreatment could decrease the mRNA levels of α-synuclein via decreasing the protein levels of IRP1. These results provide new findings and new strategies for the prevention and treatment of PD.

ZeGang Ma, Medical College of Qingdao University, China

Parkinson’s disease (PD) pathology is characterized by the formation of intra-neuronal inclusions called Lewy bodies, which are comprised of alpha-synuclein (a-syn). PD is primarily a movement disorder, but patients are known to experience memory deficit and mood disorders. In the present study, we examined the effect of hA53T mutation during development on the locomotor, anxiety-like behavior and spatial learning and memory abilities. We used the open-field test, elevated plus maze, Morris water maze and rotarod tests to evaluate hA53T transgenic mice and wild-type littermate at 3, 6, and 9 months of age. Results from open field and elevated plus maze tests show A53T mice develop age-dependent changes in locomotor activity and reduced anxiety-like behavior. Results from Morris water maze task shows no obvious differences between A53T transgenic mice and the wild type in the latency to escape on the hidden platform. In the probe test, A53T mice exhibit an unexpected improvements of learning and spatial memory abilities at 3 and 9 months, as demonstrated by a significant increase in spending more time in target quadrant on 5th and 7th days. Further studies by a reverse learning test also showed that 3 and 9-month-old transgenic mice spent more time in target quadrant compared with wild-type controls. No motor coordination impairments were observed by rotarod tests in the transgenic mice at 3, 6 and 9 months. These results indicate a possible role of the A53T a-syn mutation in enhancing spatial learning and memory abilities, suggesting that these behaviors might be disturbed in hA53T mutation PD mouse model.

Lei Chen, Qingdao University, China

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels occupy a critical position in globus pallidus pacemaking activity. Morphological studies have shown the expression of HCN channels in the globus pallidus. To investigate the in vivo effects of HCN channels in the globus pallidus, extracellular recordings and behavioral tests were performed in the present study. In normal rats, micro-pressure ejection of 0.05 mM ZD7288, the selective HCN channel blocker, decreased the frequency of spontaneous firing in 21 out of the 40 pallidal neurons. The average decrease was 50.4±5.4%. Interestingly, in another 18 out of the 40 pallidal neurons, ZD7288 increased the firing rate by 137.1±27.6%. Similar bidirectional modulation on the firing rate was observed by a higher concentration of ZD7288 (0.5 mM) as well as another HCN channel blocker, CsCl. Furthermore, activation of HCN channels by 8-Br-cAMP increased the firing rate by 63.0±9.3% in 15 out of the 25 pallidal neurons and decreased the firing rate by 46.9±9.4% in another 8 out of the 25 pallidal neurons. Further experiments revealed that modulation of glutamatergic but not GABAergic transmission may be involved in ZD7288-induced increase in firing rate. Consistent with electrophysiological results, further studies revealed that modulation of HCN channels also had bidirectional effects on behavior. Taken together, the present studies suggest that HCN channels may modulate the activity of pallidal neurons by different pathways in vivo.

Wen-Fang Chen is a professor of Medical College of Qingdao University, China. He serves as standing director of Chinese Society for Physiology, the Chinese Society for Neuroscience and the chairman of Shandong Physiological Society.

Accumulating clinical and experimental evidence suggests that chronic neuroinflammation is associated with the progressive death of dopaminergic neurons in Parkinson’s disease (PD). The G protein-coupled estrogen receptor (GPER) was reported to be a novel membrane estrogen receptor, which responds to estrogen and mediates estrogen’s rapid cellular effects. Studies have shown that activation of GPER by selective agonist, G1, could account for some of the protective effects of estrogen against inflammatory responses in animal model of multiple sclerosis. The present study aimed to evaluate the protective effects of G1 on lipopolysaccharide (LPS)-induced microglia activation. We demonstrated that GPER was expressed in BV2 microglial cells. G1 treatment significantly inhibited the LPS-induced production of proinflammatory cytokines such as iNOS，COX2、IL-1β and TNFα. These effects could be abolished by GPER antagonist G15 and lentivirus-mediated knockdown of GPER. Moreover, LPS treatment significantly increased the phosphorylation level of ERK, JNK and p38 in BV2 microglial cells. G1 pretreatment could remarkably attenuate these changes, while co-treatment with G15 could reverse the protective effects of G1. Taken together, this study demonstrated that GPER agonist G1 can prevent LPS-induced activation of microglia. GPER may be a novel therapeutic target for chronic inflammatory diseases. Acknowledgement: This work was supported by National Natural Science Foundation of China (31271150) and the State Key Program of National Natural Science of China (81430024).

Ik-Hyun Cho is a professor of Department of Convergence Medical Science, College of Oriental Medicine, Kyung Hee University, Seoul, Korea. He has published 15 papers in reputed journals and has been serving as an editorial board member of several reputed journals.

The potential neuroprotective value of sulforaphane (SFN) in Huntington’s disease (HD) has not been established. We investigated whether SFN prevents and improves the neurological impairment and striatal cell death in a 3-nitropropionic acid (3-NP)-induced mouse model of HD. SFN (2.5 and 5.0 mg/kg/day, i.p.) was given daily 30 minutes before 3-NP treatment (pretreatment) and from onset/progression/peak points of the neurological disorders. Pretreatment with SFN (5.0 mg/kg/day) produced the best neuroprotective effect for the neurological disorders and lethality among other conditions. The protective effects due to pretreatment with SFN were associated with suppression of the formation of lesion area, neuronal death, succinate dehydrogenase activity, apoptotic cell death, microglial activation, and mRNA or protein expression of inflammatory mediators including tumor necrosis factor-alpha, interleukin (IL)-1β, IL-6, inducible nitric oxide synthase, and cyclooxygenase-2 in the striatum after 3-NP treatment. Also, pretreatment with SFN activated the Kelch-like ECH associated protein 1 (Keap1) - nuclear factor erythroid 2-related factor 2 (Nrf2) – antioxidant response element (ARE) pathway and inhibited the mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB) pathways in the striatum after 3-NP treatment. As expected, the pretreatment with activators (dimethyl fumarate and antioxidant response element inducer-3) of the Keap1-Nrf2-ARE pathway decreased the neurological impairment and lethality after 3-NP treatment. Our findings suggest that SFN may effectively attenuate 3-NPinduced striatal toxicity by activating the Keap1-Nrf2-ARE pathway and inhibiting the MAPKs and NF-κB pathways and that SFN has a wide therapeutic time-window for HD-like symptoms.

Jun Xia Xie is currently a Professor of the Department of Physiology and vice president of Qingdao University. Prof. Xie is the principle of intensely-contributed subject in Physiology of Shandong Province and chairman of Center for Neuroscience Research. She was awarded the title as “prominent scientist in Shandong Province” and “top person in Qingdao City”. She also serves as standing director of Chinese Society for Physiology, the Chinese Society for Neuroscience and the chairman of Shandong Physiological Society.

Parkinson’s disease (PD) is a common neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantial nigra pars compacta (SNpc) and the appearance of fibrillar aggregates of α-synuclein called Lewy bodies (LBs). Rosmarinic acid (RA) is an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid, which is commonly found in various plant families. It has been demonstrated that RA has a number of interesting biological activities, e.g. antiviral, antibacterial, anti-inflammatory and antioxidant activities. However, the effect of RA on α-synuclein aggregation in PD models and the molecular mechanisms underlying this effect are still not elucidated. Therefore, in this study, we investigated the effects of RA against rotenone-induced α-synuclein aggregation in C57BL/6 mice and SH-SY5Y dopaminergic cells and to explore the possible mechanisms involved in this process. Results showed that mice which had orally received rotenone for 56 d showed a significantly motor dysfunction; 50 mg/kg RA pretreatment significantly improved the motor dysfunction. The number of TH-positive neurons and the expression of TH were both reduced in the SNpc of rotenone-treated mice. This could be significantly inhibited by RA pretreatment. In addition, RA pretreatment significantly inhibited rotenone-induced increase in α-synuclein expression. In vitro study showed that RA pretreatment partially restored cell loss induced by rotenone and inhibited rotenone-induced increase in α-synuclein mRNA expression in SH-SY5Y cells. The mechanisms underlying this protection might be related to the antioxidation effect of RA by inhibiting ROS generation and increasing SOD expression. These results provide new findings and new strategies for the prevention and treatment of PD.

Habata has study his PhD in Kitasato University School of Medicine. He is a Kitasato University Lecturer, Master of Engineering and an Occupational Therapist of Japan for 30 years.

Objectives: The antiparkinsonian effect of N-methyl-D-aspartate receptor subtype 2B (NR2B) antagonists remain controversial. To determine the effects of NR2B antagonist on Parkinson’s disease (PD), the effect of a single administration of D1R agonist SKF38393 (SKF) and NR2B antagonist ifenprodil or co-administration of ifenprodil and SKF were investigated using the cylinder test. Methods: We performed the cylinder test to estimate the number of forelimb use per 15 min as a measure of motor activity 30 min after administration of SKF(1.0, 2.0, 3.0 mg/kg), 0.1 mg/kg ifenprodil or co-administration of SKF and ifenprodil. Next, to identify the brain areas influenced by SKF and ifenprodil, neurons with SKF-induced c-Fos expression were analysed in various brain regions in hemi-PD following the administration of SKF, with and without ifenprodil. Results: The administration of SKF increased a frequency of forelimb-use in dose-dependent manner, mostly via the facilitation of the frequent use of parkinsonian paw. The combined administration of SKF and ifenprodil completely reversed the SKFinduced abnormality in bilaterally coordinated movement of the forelimb in hemi-PD without affecting the facilitatory effect of SKF on motor activity. The co-administered ifenprodil also modulated the SKF-induced c-Fos expression in the striatum and the subthalamic nucleus (STN). Conclusion: We conclude that the ameliorative effect of ifenprodil on the motor deficits in hemi-PD is resulted from the improvement of the SKF-induced excessive use of parkinsonian paw, and that the STN and/or the striatum in the lesioned hemisphere are possible targets for the antiparkinsonian effect of the NR2B antagonist.

Sanjay Jaiswal has completed his MD from RNT Medical College, Udaipur, India and DM Neurology from Institute of Medical Sciences BHU, Varanasi, India. He is a Consultant Neurologist at Jaiswal Hospital and Neuro Institute, Kota, Rajasthan, India. He has published several papers in reputed journals and has been serving as a Faculty in most of the neurology conferences. He has got an extensive experience of organizing and conducting conferences in the field of neurology and has been awarded by State Government for the contribution to social welfare.

Dystonia, defined as a neurological syndrome characterised by involuntary, patterned, sustained, or repetitive muscle contractions of opposing muscles, causing twisting movements and abnormal postures, is one of the most disabling movement disorders. A small portion of patients have a known cause and respond to specific treatments, such as levodopa in dopa-responsive dystonia or drugs that prevent copper accumulation in Wilson’s disease. Therapeutic options must be tailored to the needs of individual patients. In patients on chronic dopaminergic treatment, peak-dose dystonia, diphasic dystonia and off-dystonia can be seen. The later constitutes the major dystonic feature of chronic levodopa therapy, and a wide variety of strategies are available to manage this complication. Many therapeutic options are available viz. Chemo-denervation with botulinum toxin injections and medical treatments. Among them, Deep Brain Stimulation (DBS) of the sub-thalamic nucleus has proved to be the most effective one. Parkinsonian tremor is most likely due to oscillating neuronal activity within the CNS. The most likely candidate producing these oscillations is the basal ganglia loop and its topographic organization might be responsible for the separation into different oscillators which, nevertheless, usually produce the same frequency. The relative efficacy of trihexiphenidyl hydrochloride, amantadine hydrochloride, and low-dose carbidopa-levodopa in reducing Parkinsonian tremors was investigated using objective techniques. Trihexiphenidyl and carbidopa-levodopa decreased tremor by greater than 50%. Some patients responded to one drug but not to the other. Amantadine decreased tremor less than 25%. Monotherapy with trihexiphenidyl or carbidopa-levodopa should be the initial treatment for the tremor of Parkinson’s disease.

Naik Bakht Arbabzada, research assistant for Dr. Keir Pearson in University of Alberta, Canada. Research Assistant for Dr. Ian Winship Alberta's Health Research and Innovation. Section rat brain and carryout different studies (immunohistochemistry and staining) on the slides and analyzed the tissue for different protein markers.

The long acting Beta2 Adrenergic Receptor (β2AR) Agonist salmeterol has been shown to be neuro‐ protective against the loss of dopaminergic neurons in animal models of Parkinson’s disease (PD). Salmeterol was found to work by inhibiting the inflammatory response of microglial cells, a key cell in the pathogenesis of PD. Recently, super-long acting β2AR agonists, vilanterol andindacaterol, have been described, and the objective of this study was to examine the effects of these super longacting agonists on the inflammatory response of the microglial cell line BV-2, and compare their mode of action to that of salmeterol. We found that the dose-dependent inhibitory action of both indacaterol and vilanterol are similar to that of salmeterol with respects to the production of TNF-alpha by BV-2 cells stimulated with the inflammogen LPS. Conversely, we find that at higher concentrations indacaterol, vilanterol and salmeterol enhance IL-6 production and release by stimulated BV-2 cells, while lower concentrations exert an inhibitory effect on the IL-6 production and release.Furthermore, like salmeterol, indacaterol and vilanterol also exert their inhibitory effecton TNF-α production through the inhibition of NF-kB in a TAK-1 and Beta-arrestin-2-dependent manner. Interestingly, both these b2AR agonists inhibit the classical and alternate pathways of NF-kB, but the kinetics of inhibition vary between pathways. These findings provide further insight into how β2AR agonists work to reverse the CNS inflammation that occurs in Parkinson disease, and may provide a new and more effective therapeutic for PD.

Anwar ullah is enrolled as a PhD student at Quaid-i-Azam University Islamabad. Currently he is working as a visiting scholar with synaptic signaling and neurodegeneration group at German center for neurodegenerative diseases (DZNE). His work includes investigating genetic causes of familial and sporadic Parkinson’s disease in Pakistani population.

CYP2D6 gene (OMIM: 124030) encodes a member of the cytochrome P450 superfamily of enzymes involved in metabolism of several psychoactive drugs as well as in the neurotransmitter biotransformation in brain. Allelic variants at CYP2D6 locus underlie altered dopamine metabolism pathway and therefore considered to be as a possible genetic factor contributing to Parkinson’s disease (PD) susceptibility. Population based genetic epidemiological studies exploring the association between a most common null polymorphic variant of CYP2D6 (CYP2D6*4; 1846G > A) and PD risk have revealed controversial results. The role of CYP2D6 null alleles as a risk factor for PD therefore remains elusive.. In the present study we used an association approach to estimate the contribution of CYP2D6*4 allelic polymorphism (1846G > A) on the risk of PD in Pakistani population. Allele specific PCR assay targeting 1846G > A polymorphism was performed in 174 clinically characterized unrelated PD patients and 200 age, gender and ethnicity-matched controls. No deviation of genotype distribution from Hardy-Weinberg equilibrium was found in the control group (X2=0.44, P = 0.507). There was however a significant difference of genotype and allele frequencies between cases and controls at CYP2D6 locus with respect to 1846G > A polymorphism. Minor allele (A) encoding functionally defective form of CYP2D6 enzyme (CYP2D6*4) increased the risk for PD more than two fold (OR =2.525; 95% CI =1.410-4.522; P = 0.00135). A comparison of the genotype frequencies for homozygous minor allele (AA-genotype) revealed a further 10-fold increase in the risk of PD (OR: 21.32; 95% CI: 1.22 - 372.5). Our results demonstrate an increased in minor allele and genotype frequencies for CYP2D6 (1846G > A) polymorphism in Pakistani samples that implies clinical significance of CYP2D6 locus in this population.