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FP48-TH-04 Cerebrospinal fluid biomarkers and their dysregulation in amyotrophic lateral sclerosis
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19th World Congress of Neurology, Free Paper Abstracts / Journal of the Neurological Sciences 285 S1 (2009) S57–S154
FP48 – Neuroimmunology FP48-TH-01 Immunopathological study of pseudohypertrophy of the inferior olivary nucleus K. Ogawa1 , T. Mizutani1 , K. Uehara1 , M. Minami1 , Y. Suzuki2 , T. Uchihara3 . 1 Division of Neurology, Department of Medicine, Nihon University School of Medicine, Tokyo, Japan; 2 Department of Pathology, Asahi General Hospital, Chiba, Japan; 3 Department of Neurology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan There have been only a few reports about the immunohistochemical study of pseudohypertrophy of the inferior olivary nucleus (PHIO). We performed the detailed immunohistochemical study of 10 PH-IOs in 8 patients to clarify the mechanism of neuronal degeneration of PH-IO. We used antibodies to aB-crystallin (aBC), synaptophysin (SYP), microtubule-associated protein 2 (MAP2), LysAsp-Glu-Leu (KDEL) receptors, heat shock protein (HSP) 27 as well as SMI-31. We found aBC-positive neurons on the ipsilateral side of 10 PH-IOs. SMI-31-positive neurons were also observed in 6 PH-IOs. Confocal laser microscopy showed co-localization of aBC and SMI-31 in some neurons in PH-IO. However, there were no HSP27-positive neurons or astrocytes in any of the 10 PH-IOs. MAP2 immunostaining showed MAP2-positive hypertrophic thick neurites around hypertrophic neurons on the ipsilateral side of 7 PH-IOs and demonstrated glomeruloid structures in 3 PH-IOs. In addition, fine granular SYP-immunoreactivity was decreased in the neuropils on the ipsilateral side of all 10 PH-IOs. SYPimmunoreactive dots were scattered in the neuropils and on the neuronal cell bodies on the side of 7 PH-IOs, and the aggregation of SYP-immunoreactive dots scattered in the neuropils was shown in 3 PH-IOs. Double-immunostainings using anti-MAP2 and anti-SYP antibodies demonstrated frequent SYP-immunoreactive dots along the MAP2-positive hypertrophic thick neurites and their cell bodies. Periphery-stained KDEL-positive neurons were also found on the side of 7 PH-IOs. We showed that the change of the distribution of presynaptic terminals correlated well to the hypertrophic thick neurites in PH-IO. aBC is one of the small heat shock proteins. We considered that aBC was expressed in the neurons in PH-IO, induced by cellular stress. KDEL receptors of the neurons in PH-IO were distributed to the periphery of the cytoplasm. This change of the distribution of KDEL receptors was considered to be due to PH-IO. FP48-TH-02 Using intravenous immunoglobulin in the treatment of neurological diseases: a review of 40 cases and analysis of EFNS guideline orientations H.M.G. Martins1 , M. Neves1 , A.V. Herrero2 . 1 Servico ¸ de Medicina I, Hospital Prof. Dr. Fernando Fonseca, Amadora, Portugal; 2 Servico ¸ de Neurologia, Hospital Prof. Dr. Fernando Fonseca, Amadora, Portugal Purpose: Intravenous immunoglobulin (IVIG) is increasingly used to treat various neurologic diseases. EFNS recommendations are stringent but have helped to clarify its use. The aim of this study was to review real uses of IVIG in a neurology department contrasting to suggested recommendations. Methods: In a retrospective study of all hospitalizations in neurology where IVIG treatment was performed 40 cases were found. Variables considered were: age, gender, diagnosis (according to EFNS guideline), clinical improvement, number of subsequent cycles and whether EFNS recommendations were followed or not. Data was analyzed using Excel® , EpiInfo® software. A 95% CI, Qui2 and Fisher exact tests were used. Results: Average age was 55.7 y [21–84 y] and 63% were males. List of conditions: GBS 12, CIDP 8, MG acute exacerbation 6, others 14 including: Devic S, IBM (2), LEMS (2), MFS (3), MG, MMN, PM/IBM, PPS, Myeloradiculitis, Probable Romboencefalomielitis of Bickerstaff. EFNS recommendations were Followed (78%) Not
followed (20%) and absent in 1 case of Myeloradiculitis. Clinical outcomes were: none 20%, not significant 10%, modest 27.5%, very significant 42.5%. Due to good response 14 patients repeated IVIG treatment, 7 (1 time), 3 (2 times), 4 patients repeated: 3, 4, 10 and 15 times. No IVIG side effects were reported. Positive outcomes (Modest+Very Significant) were associated with following EFNS guidelines (OR = 8.3, CI 1.25–62.5, p = 0.012). From the 31 cases where IVIG was EFNS recommended, there was significant clinical improvement in 25 and none to little improvement in 6. From 9 cases without recommendation for treatment, 1/3 actually improved significantly (two cases of Miller Fisher syndrome and one of Inclusion Body Miositis). Conclusions: Treatment according to EFNS guidelines correlates strongly with success, which suggests these guidelines are useful in clinical practice. In a limited number of cases not following the guidelines, however, clinical outcomes were favorable. FP48-TH-03 Proinflammatory cytokines in different types of dementia N. Lobjanidze, R. Shakarishvili, M. Beridze, M. Janelidze, E. Melikidze. Neurology, State Medical University Clinic, Tbilisi, Georgia Aim: Study aimed at investigation of the effect of IL-1, IL-6, TNF-a blood plasma expression on progression of cognitive decline in patients with Dementia. Subjects and Methods: Thirty demented patients aged 50 to 75, 15 patients with VAD (Vascular Dementia) and 15-with AD (Alzheimer’s disease) have been investigated prospectively for 18months. Control comprised 15 age-matched healthy individuals. The NINDS-AIREN criteria and diagnostic tool ICD-10 were used to establish diagnosis of VAD. The NINDS-ADRDA criteria used for diagnostics of AD. Neuroimaging studies performed by conventional MRI. Blood immunochemical substrates (IL-1b, IL-6, TNF-a) were researched prospectively 3 times every 6 months using the ELISA method. Cognitive functions determined accordingly by MMSE and Wechsler memory scale in VAD and AD and control. Results: Blood proinflammatory cytokines in AD and VAD patients were elevated against control (p < 0.05). In AD patients the level of TNF-a was significantly increased compared to VAD patients. Significant prospective changes were not found in levels of IL- 1b, IL-6, TNF-a in VAD patients, while the levels of IL-6 were elevated after 18 months period in AD patients. Cognition did not change significantly in VAD patients during 18 months period. Correlation was found between the blood IL-6 levels and Memory decline in AD patients (r= + 0.21, p < 0.05). Conclusion: Probably, inflammatory molecules play the key role in development of cognitive decline in AD. FP48-TH-04 Cerebrospinal fluid biomarkers and their dysregulation in amyotrophic lateral sclerosis T. Tateishi, R. Yamasaki, M. Tanaka, M. Fukunaga, H. Kikuchi, T. Matsushita, K. Motomura, Y. Ohyagi, J. Kira. Neurology, Kyushu University, Fukuoka, Japan Purpose: To identify biomarkers related to amyotrophic lateral sclerosis (ALS) pathogenesis. Method: We measured the levels of 27 cytokines/chemokines and growth factors in cerebrospinal fluid from 42 patients with sporadic ALS, 12 patients with lower motor neuron disease, and 34 control patients, using a multiplexed fluorescent bead-based immunoassay. We then performed densitometric analyses focusing on members of the vascular endothelial growth factor (VEGF) pathway in six postmortem ALS cases and six non-neurological disease controls, together with G93A mutant superoxide dismutase 1 (mSOD1) transgenic ALS model mice. Results: Levels of VEGF, granulocyte-colony stimulating factor, CCL2, CCL4, CXCL8, CXCL10, and several other proinflammatory
19th World Congress of Neurology, Free Paper Abstracts / Journal of the Neurological Sciences 285 S1 (2009) S57–S154
cytokines were significantly higher in ALS patients than controls. CCL2 levels were negatively correlated with the revised ALS functional rating scale score, while CCL4 and VEGF showed positive correlations with the score. CCL4 and CXCL10 showed negative correlations with disease progression rate. Immunoreactivities for VEGF and VEGF receptors 1 and 2 in anterior horn cells were significantly weaker in ALS cases than controls, whereas hypoxiainducible factor (HIF)-1a immunoreactivity in anterior horn cells was significantly stronger in ALS cases. VEGF immunoreactivity in anterior horn cells was weaker in mSOD1 transgenic mice than in non-transgenic mice aged from 12 (presymptomatic stage) to 18 weeks (end stage), despite the stronger HIF-1a-immunoreactivity in mSOD1 transgenic mice than in non-transgenic mice. Conclusion: These findings suggest that CCL2 is a neurotoxic biomarker, that CCL4 and CXCL10 are neuroprotective markers, and that blockade of the HIF-1a/VEGF pathway together with down-modulation of VEGF receptors in motor neurons contribute to their death in ALS, even though up-regulated proinflammatory cytokines/chemokines augment VEGF secretion in astrocytes. FP48-TH-05 Study of low dose naltrexone modulation of the endocannabinoid system in patients with multiple sclerosis M. Gironi1 , N. Pasquariello2 , S. Franchi3 , P. Sacerdote3 , G. Martino4 , F. Martinelli-Boneschi4 , V. Martinelli4 , G. Comi4 , R. Nemni1 , C. Solaro5 , D. Centonze6 , M. Maccarone2 . 1 Don Gnocchi Foundation, IRCCS, Milano, Italy; 2 Department of Biomedic Sciences, Teramo, Italy; 3 Department of Pharmacology, Milano, Italy; 4 INSPE, Fondazione San Raffaele, Milano, Italy; 5 Ospedale Micone, Genova, Italy; 6 Department Of Neurosciences, Rome, Italy Introduction: The use of low dose naltrexone (LDN) in Multiple Sclerosis (MS) largely relies on anecdotal reports. We recently showed that LDN (4 mg/die) administered daily for 6 months in 40 patients with primary progressive (PP) MS (Gironi et al., 2008) was safe, well tolerated and partially efficacious on spasticity. Therapeutic efficacy was associated with a prolonged release of endogenous opioids such as Beta-endorphin (BE) from peripheral mononuclear cells (PBL). Considering the extensively reported interaction between opioids and the endocannabinoid system (ECS) we investigated the expression of anandamide-binding type-1 and type-2 cannabinoid receptors (CB1R and CB2R), as well as of NAPEPLD and FAAH, in 13 PP-MS patients treated daily for 6 months with LDN. Fifteen healthy subjects (HC) were used as control group. Methods: Quantitative real-time PCR for CB1R and CB2R, NAPE and FAHH was performed before (time 0), after 6 months of LDN therapy, and one month after drug discontinuation in PBL. Actin was used as housekeeping gene. RESULTS: CB1R and CB2R expression levels were 18-fold and 21-fold increased in MS compared to HC, respectively (p < 0.001). CB1R and CB2R decreased after 6 months of LDN therapy, altough CB1R tended to increase to normal values one month after drug discontinuation. NAPE (23-fold increase) and FAAH (7-fold increase) levels were higher in PP-MS compared to HC (p < 0.01) irrespective of the time point of measurement. Conclusions: Our data show that LDN treatment might reduce the expression of anandamide-metabolizing enzymes and anandamidebinding receptors. This observation is noteworthy, because it suggests that whatever the molecular details of ECS dysregulation, the drug is able to alleviate it. Moreover it suggest a a biological activity of LDN in MS patients.
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FP49 – Dementia: non-Alzheimer’s dementia and behavior neurology FP49-TH-01 Behavioural changes and cognitive impairment in motor neurone disease P. Lillo, S. Savage, S. Hsieh, E. Mioshi, C. Leyton, J.R. Hodges. Frontotemporal Dementia Group, Prince of Wales Medical Research Institute, Sydney, Australia Purpose: To identify cognitive and behavioural changes in patients with motor neurone disease (MND). To assess the potential impact of these changes on carer burden. Methods: A postal survey was completed by patients with MND (n = 83) and their carers (n = 75) from New South Wales. The survey covered basic demographic information, motor symptoms and neuropsychiatric symptoms in the patients. Carers received questionnaires regarding their current mood and burden of care. In addition, a subset of 13 participants undertook a cognitive screening following referral from a specialist MND clinic. Results were compared with age and gender matched controls. Results: Carers reported severe behavioural changes in MND patients involving principally motivation (45%), but also mood, stereotypic/motor behaviours and abnormal behaviour. Less than 25% of the carers reported depression but 44% reported severe burden. The carer burden was explained predominantly by abnormal behaviour and impairment of everyday skills in MND patients. Patients with MND presenting behavioural changes performed worse on the cognitive screening (Addenbroke’s Cognitive Examination Revised/ACE-R) in memory, fluency and language than controls (p < 0.01). 7 of 13 MND patients presented scores below the cut off (82) indicating significant cognitive impairment. Discussion: The study suggested a high prevalence of behavioural changes in MND patients principally in motivation. Behavioural changes in patients with MND had a greater impact on carer burden than the physical disability. Patients with behavioural changes tended to present a worse performance on cognitive testing. FP49-TH-02 The importance of nicotine in a 6-hydroxydopamine-induced rat model of Parkinson’s disease A.S.U. Ciobica1 , L.G. Hritcu1 , V. Artenie1 , W.U. Bild2 . 1 Alexandru Ioan Cuza University, Iasi, Romania; 2 Romanian Academy, Iasi, Romania Introduction: The most widely used animal models of Parkinson’s disease involve intracranial infusion of the neurotoxin 6-hydroxydopamine (6-OHDA) directly into the ascending dopaminergic forebrain bundle, thereby, inducing severe dopaminergic neuronal degeneration associated with profound deficits in cognitive functions. Aim: The aim of the present work was to study the effects of right-unilateral 6-OHDA lesions of the ventral tegmental area (VTA) or substantia nigra pars reticulata (SNr) on learning and memory processes evidenced by means of Y-maze task and shuttle-box task, respectively. We also examined the effect of nicotine treatment on the 6-OHDA lesioned rats. Our data suggest that a correlation exist between VTA, SNr and nAchRS and expression of cognitive capacities. Method: Male Wistar rats were subjected to right-unilateral 6-hydroxydopamine (6-OHDA) lesion of the ventral tegmental area or substantia nigra pars reticulata, or were sham lesioned, and nicotine treatment and their ability to acquire the operant task was studied by means of Y-maze task and shuttle-box task, respectively. The sham rats were injected with saline. Learning and memory tests were started 2 weeks after the operation. Results: Lesions of both the VTA and SNr resulted in an impairment of both conditioned avoidance response and crossing latency tested by means of shuttle-box task, suggesting significant effects of
19th World Congress of Neurology, Free Paper Abstracts / Journal of the Neurological Sciences 285 S1 (2009) S57–S154
FP48 – Neuroimmunology FP48-TH-01 Immunopathological study of pseudohypertrophy of the inferior olivary nucleus K. Ogawa1 , T. Mizutani1 , K. Uehara1 , M. Minami1 , Y. Suzuki2 , T. Uchihara3 . 1 Division of Neurology, Department of Medicine, Nihon University School of Medicine, Tokyo, Japan; 2 Department of Pathology, Asahi General Hospital, Chiba, Japan; 3 Department of Neurology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan There have been only a few reports about the immunohistochemical study of pseudohypertrophy of the inferior olivary nucleus (PHIO). We performed the detailed immunohistochemical study of 10 PH-IOs in 8 patients to clarify the mechanism of neuronal degeneration of PH-IO. We used antibodies to aB-crystallin (aBC), synaptophysin (SYP), microtubule-associated protein 2 (MAP2), LysAsp-Glu-Leu (KDEL) receptors, heat shock protein (HSP) 27 as well as SMI-31. We found aBC-positive neurons on the ipsilateral side of 10 PH-IOs. SMI-31-positive neurons were also observed in 6 PH-IOs. Confocal laser microscopy showed co-localization of aBC and SMI-31 in some neurons in PH-IO. However, there were no HSP27-positive neurons or astrocytes in any of the 10 PH-IOs. MAP2 immunostaining showed MAP2-positive hypertrophic thick neurites around hypertrophic neurons on the ipsilateral side of 7 PH-IOs and demonstrated glomeruloid structures in 3 PH-IOs. In addition, fine granular SYP-immunoreactivity was decreased in the neuropils on the ipsilateral side of all 10 PH-IOs. SYPimmunoreactive dots were scattered in the neuropils and on the neuronal cell bodies on the side of 7 PH-IOs, and the aggregation of SYP-immunoreactive dots scattered in the neuropils was shown in 3 PH-IOs. Double-immunostainings using anti-MAP2 and anti-SYP antibodies demonstrated frequent SYP-immunoreactive dots along the MAP2-positive hypertrophic thick neurites and their cell bodies. Periphery-stained KDEL-positive neurons were also found on the side of 7 PH-IOs. We showed that the change of the distribution of presynaptic terminals correlated well to the hypertrophic thick neurites in PH-IO. aBC is one of the small heat shock proteins. We considered that aBC was expressed in the neurons in PH-IO, induced by cellular stress. KDEL receptors of the neurons in PH-IO were distributed to the periphery of the cytoplasm. This change of the distribution of KDEL receptors was considered to be due to PH-IO. FP48-TH-02 Using intravenous immunoglobulin in the treatment of neurological diseases: a review of 40 cases and analysis of EFNS guideline orientations H.M.G. Martins1 , M. Neves1 , A.V. Herrero2 . 1 Servico ¸ de Medicina I, Hospital Prof. Dr. Fernando Fonseca, Amadora, Portugal; 2 Servico ¸ de Neurologia, Hospital Prof. Dr. Fernando Fonseca, Amadora, Portugal Purpose: Intravenous immunoglobulin (IVIG) is increasingly used to treat various neurologic diseases. EFNS recommendations are stringent but have helped to clarify its use. The aim of this study was to review real uses of IVIG in a neurology department contrasting to suggested recommendations. Methods: In a retrospective study of all hospitalizations in neurology where IVIG treatment was performed 40 cases were found. Variables considered were: age, gender, diagnosis (according to EFNS guideline), clinical improvement, number of subsequent cycles and whether EFNS recommendations were followed or not. Data was analyzed using Excel® , EpiInfo® software. A 95% CI, Qui2 and Fisher exact tests were used. Results: Average age was 55.7 y [21–84 y] and 63% were males. List of conditions: GBS 12, CIDP 8, MG acute exacerbation 6, others 14 including: Devic S, IBM (2), LEMS (2), MFS (3), MG, MMN, PM/IBM, PPS, Myeloradiculitis, Probable Romboencefalomielitis of Bickerstaff. EFNS recommendations were Followed (78%) Not
followed (20%) and absent in 1 case of Myeloradiculitis. Clinical outcomes were: none 20%, not significant 10%, modest 27.5%, very significant 42.5%. Due to good response 14 patients repeated IVIG treatment, 7 (1 time), 3 (2 times), 4 patients repeated: 3, 4, 10 and 15 times. No IVIG side effects were reported. Positive outcomes (Modest+Very Significant) were associated with following EFNS guidelines (OR = 8.3, CI 1.25–62.5, p = 0.012). From the 31 cases where IVIG was EFNS recommended, there was significant clinical improvement in 25 and none to little improvement in 6. From 9 cases without recommendation for treatment, 1/3 actually improved significantly (two cases of Miller Fisher syndrome and one of Inclusion Body Miositis). Conclusions: Treatment according to EFNS guidelines correlates strongly with success, which suggests these guidelines are useful in clinical practice. In a limited number of cases not following the guidelines, however, clinical outcomes were favorable. FP48-TH-03 Proinflammatory cytokines in different types of dementia N. Lobjanidze, R. Shakarishvili, M. Beridze, M. Janelidze, E. Melikidze. Neurology, State Medical University Clinic, Tbilisi, Georgia Aim: Study aimed at investigation of the effect of IL-1, IL-6, TNF-a blood plasma expression on progression of cognitive decline in patients with Dementia. Subjects and Methods: Thirty demented patients aged 50 to 75, 15 patients with VAD (Vascular Dementia) and 15-with AD (Alzheimer’s disease) have been investigated prospectively for 18months. Control comprised 15 age-matched healthy individuals. The NINDS-AIREN criteria and diagnostic tool ICD-10 were used to establish diagnosis of VAD. The NINDS-ADRDA criteria used for diagnostics of AD. Neuroimaging studies performed by conventional MRI. Blood immunochemical substrates (IL-1b, IL-6, TNF-a) were researched prospectively 3 times every 6 months using the ELISA method. Cognitive functions determined accordingly by MMSE and Wechsler memory scale in VAD and AD and control. Results: Blood proinflammatory cytokines in AD and VAD patients were elevated against control (p < 0.05). In AD patients the level of TNF-a was significantly increased compared to VAD patients. Significant prospective changes were not found in levels of IL- 1b, IL-6, TNF-a in VAD patients, while the levels of IL-6 were elevated after 18 months period in AD patients. Cognition did not change significantly in VAD patients during 18 months period. Correlation was found between the blood IL-6 levels and Memory decline in AD patients (r= + 0.21, p < 0.05). Conclusion: Probably, inflammatory molecules play the key role in development of cognitive decline in AD. FP48-TH-04 Cerebrospinal fluid biomarkers and their dysregulation in amyotrophic lateral sclerosis T. Tateishi, R. Yamasaki, M. Tanaka, M. Fukunaga, H. Kikuchi, T. Matsushita, K. Motomura, Y. Ohyagi, J. Kira. Neurology, Kyushu University, Fukuoka, Japan Purpose: To identify biomarkers related to amyotrophic lateral sclerosis (ALS) pathogenesis. Method: We measured the levels of 27 cytokines/chemokines and growth factors in cerebrospinal fluid from 42 patients with sporadic ALS, 12 patients with lower motor neuron disease, and 34 control patients, using a multiplexed fluorescent bead-based immunoassay. We then performed densitometric analyses focusing on members of the vascular endothelial growth factor (VEGF) pathway in six postmortem ALS cases and six non-neurological disease controls, together with G93A mutant superoxide dismutase 1 (mSOD1) transgenic ALS model mice. Results: Levels of VEGF, granulocyte-colony stimulating factor, CCL2, CCL4, CXCL8, CXCL10, and several other proinflammatory
19th World Congress of Neurology, Free Paper Abstracts / Journal of the Neurological Sciences 285 S1 (2009) S57–S154
cytokines were significantly higher in ALS patients than controls. CCL2 levels were negatively correlated with the revised ALS functional rating scale score, while CCL4 and VEGF showed positive correlations with the score. CCL4 and CXCL10 showed negative correlations with disease progression rate. Immunoreactivities for VEGF and VEGF receptors 1 and 2 in anterior horn cells were significantly weaker in ALS cases than controls, whereas hypoxiainducible factor (HIF)-1a immunoreactivity in anterior horn cells was significantly stronger in ALS cases. VEGF immunoreactivity in anterior horn cells was weaker in mSOD1 transgenic mice than in non-transgenic mice aged from 12 (presymptomatic stage) to 18 weeks (end stage), despite the stronger HIF-1a-immunoreactivity in mSOD1 transgenic mice than in non-transgenic mice. Conclusion: These findings suggest that CCL2 is a neurotoxic biomarker, that CCL4 and CXCL10 are neuroprotective markers, and that blockade of the HIF-1a/VEGF pathway together with down-modulation of VEGF receptors in motor neurons contribute to their death in ALS, even though up-regulated proinflammatory cytokines/chemokines augment VEGF secretion in astrocytes. FP48-TH-05 Study of low dose naltrexone modulation of the endocannabinoid system in patients with multiple sclerosis M. Gironi1 , N. Pasquariello2 , S. Franchi3 , P. Sacerdote3 , G. Martino4 , F. Martinelli-Boneschi4 , V. Martinelli4 , G. Comi4 , R. Nemni1 , C. Solaro5 , D. Centonze6 , M. Maccarone2 . 1 Don Gnocchi Foundation, IRCCS, Milano, Italy; 2 Department of Biomedic Sciences, Teramo, Italy; 3 Department of Pharmacology, Milano, Italy; 4 INSPE, Fondazione San Raffaele, Milano, Italy; 5 Ospedale Micone, Genova, Italy; 6 Department Of Neurosciences, Rome, Italy Introduction: The use of low dose naltrexone (LDN) in Multiple Sclerosis (MS) largely relies on anecdotal reports. We recently showed that LDN (4 mg/die) administered daily for 6 months in 40 patients with primary progressive (PP) MS (Gironi et al., 2008) was safe, well tolerated and partially efficacious on spasticity. Therapeutic efficacy was associated with a prolonged release of endogenous opioids such as Beta-endorphin (BE) from peripheral mononuclear cells (PBL). Considering the extensively reported interaction between opioids and the endocannabinoid system (ECS) we investigated the expression of anandamide-binding type-1 and type-2 cannabinoid receptors (CB1R and CB2R), as well as of NAPEPLD and FAAH, in 13 PP-MS patients treated daily for 6 months with LDN. Fifteen healthy subjects (HC) were used as control group. Methods: Quantitative real-time PCR for CB1R and CB2R, NAPE and FAHH was performed before (time 0), after 6 months of LDN therapy, and one month after drug discontinuation in PBL. Actin was used as housekeeping gene. RESULTS: CB1R and CB2R expression levels were 18-fold and 21-fold increased in MS compared to HC, respectively (p < 0.001). CB1R and CB2R decreased after 6 months of LDN therapy, altough CB1R tended to increase to normal values one month after drug discontinuation. NAPE (23-fold increase) and FAAH (7-fold increase) levels were higher in PP-MS compared to HC (p < 0.01) irrespective of the time point of measurement. Conclusions: Our data show that LDN treatment might reduce the expression of anandamide-metabolizing enzymes and anandamidebinding receptors. This observation is noteworthy, because it suggests that whatever the molecular details of ECS dysregulation, the drug is able to alleviate it. Moreover it suggest a a biological activity of LDN in MS patients.
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FP49 – Dementia: non-Alzheimer’s dementia and behavior neurology FP49-TH-01 Behavioural changes and cognitive impairment in motor neurone disease P. Lillo, S. Savage, S. Hsieh, E. Mioshi, C. Leyton, J.R. Hodges. Frontotemporal Dementia Group, Prince of Wales Medical Research Institute, Sydney, Australia Purpose: To identify cognitive and behavioural changes in patients with motor neurone disease (MND). To assess the potential impact of these changes on carer burden. Methods: A postal survey was completed by patients with MND (n = 83) and their carers (n = 75) from New South Wales. The survey covered basic demographic information, motor symptoms and neuropsychiatric symptoms in the patients. Carers received questionnaires regarding their current mood and burden of care. In addition, a subset of 13 participants undertook a cognitive screening following referral from a specialist MND clinic. Results were compared with age and gender matched controls. Results: Carers reported severe behavioural changes in MND patients involving principally motivation (45%), but also mood, stereotypic/motor behaviours and abnormal behaviour. Less than 25% of the carers reported depression but 44% reported severe burden. The carer burden was explained predominantly by abnormal behaviour and impairment of everyday skills in MND patients. Patients with MND presenting behavioural changes performed worse on the cognitive screening (Addenbroke’s Cognitive Examination Revised/ACE-R) in memory, fluency and language than controls (p < 0.01). 7 of 13 MND patients presented scores below the cut off (82) indicating significant cognitive impairment. Discussion: The study suggested a high prevalence of behavioural changes in MND patients principally in motivation. Behavioural changes in patients with MND had a greater impact on carer burden than the physical disability. Patients with behavioural changes tended to present a worse performance on cognitive testing. FP49-TH-02 The importance of nicotine in a 6-hydroxydopamine-induced rat model of Parkinson’s disease A.S.U. Ciobica1 , L.G. Hritcu1 , V. Artenie1 , W.U. Bild2 . 1 Alexandru Ioan Cuza University, Iasi, Romania; 2 Romanian Academy, Iasi, Romania Introduction: The most widely used animal models of Parkinson’s disease involve intracranial infusion of the neurotoxin 6-hydroxydopamine (6-OHDA) directly into the ascending dopaminergic forebrain bundle, thereby, inducing severe dopaminergic neuronal degeneration associated with profound deficits in cognitive functions. Aim: The aim of the present work was to study the effects of right-unilateral 6-OHDA lesions of the ventral tegmental area (VTA) or substantia nigra pars reticulata (SNr) on learning and memory processes evidenced by means of Y-maze task and shuttle-box task, respectively. We also examined the effect of nicotine treatment on the 6-OHDA lesioned rats. Our data suggest that a correlation exist between VTA, SNr and nAchRS and expression of cognitive capacities. Method: Male Wistar rats were subjected to right-unilateral 6-hydroxydopamine (6-OHDA) lesion of the ventral tegmental area or substantia nigra pars reticulata, or were sham lesioned, and nicotine treatment and their ability to acquire the operant task was studied by means of Y-maze task and shuttle-box task, respectively. The sham rats were injected with saline. Learning and memory tests were started 2 weeks after the operation. Results: Lesions of both the VTA and SNr resulted in an impairment of both conditioned avoidance response and crossing latency tested by means of shuttle-box task, suggesting significant effects of