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The comparison of mesenchymal stem cell with Mitoxantrone in treatment of patients with multiple sclerosis
Abstracts
sleep disorder was the initial symptom in four patients and in the rest it was preceded by gait instability. Patients developed variable degrees of chorea, dysautonomia, dysarthria, dysphagia, central hypoventilation. All four patients tested had HLA-DRB1*1001 and HLA-DQB1*0501 alleles. All patients' serum/CSF showed the same pattern of rat brain neuropil immunostaining that could be abrogated by immunocompetition assays suggesting a similar epitope, and they also labeled the cell membrane of live neurons. Patients' antibodies were IgG4. The immunoprecipitated target antigen was IgLON5, a neuronal adhesion molecule that is part of the immunoglobulin superfamily. The IgLON family of proteins is involved in the control of neuronal pathfinding and synaptogenesis. Patients had only IgLON5 antibodies and they did not have any additional antibodies against other neuronal surface antigens or IgLON family members. One of the 298 controls who had progressive supranuclear palsy had IgLON5 antibodies in serum but not in CSF. The neuropathological examination in two patients showed a neuronal tauopathy mainly involving the hypothalamus and tegmentum of the brainstem without inflammation. IgLON5 antibodies identify a novel sleep disorder characterized by NREM and REM parasomnia and stridor with sleep apnea with pathological findings suggesting a novel tauopathy. These findings may provide a link between autoimmunity and neurodegeneration.
doi:10.1016/j.jneuroim.2014.08.118
58 Myasthenia gravis associated with thymoma in a patient with CHRNE congenital myasthenic syndrome Ernestina Santosa, Isabel Moreirab, Maria Isabel Leitec, Jose Lopes Limab a Neurology Department, Hospital Santo Antonio, Instituto de Ciencias Biomedicas Abel Salazar, Universidade do Porto, Porto, Portugal; bNeurology Department, Hospital Santo Antonio, Porto, Portugal; cDepartment of Clinical Neurology, Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom
Background: The dysfunction or loss of acetylcholine receptors (AChR) in the neuromuscular junction may have different causes. More frequently the cause is autoimmune (myasthenia gravis, MG) — which is associated with antibodies to AChR. Less commonly the disease is genetic (congenital myasthenic syndromes; CMS). Case Report: Two brothers, with non-consanguineous healthy parents, have, since the age of 4 and 6 years, ophthalmoplegia, dysphonia, fatigable difficulty in chewing and fatigable weakness in the limb muscles. Electromyography showed significant decrement in the amplitude of CMAP with repetitive stimulation, suggestive of pos-synaptic defect of neuromuscular transmission. Muscle enzymes were normal. Muscle biopsy performed in one of the brothers showed subtle abnormalities suggestive of myopathic origin. Genetic study revealed a mutation on the gene CHRNE, confirming the diagnosis of congenital myasthenic syndrome in both children. They started treatment with pyridostigmine with improvement. One of the brothers was stable until the age of 34 (2003). At that time he developed dyspnoea and worsening of the weakness in the limbs. Chest CT scan showed a mediastinal mass and several adenopathies. Thymus was removed, and the histology revealed a thymoma. The adenopathies were found to be of a thyroid papillary carcinoma. He was submitted also to thyroidectomy followed by radiotherapy. Some months later he improved returning to his baseline. Antibodies anti-AChR's were not searched during the period of the worsening and previously to the thymectomy. In 2012 they were negative.
45
Comments: These cases demonstrate that, although very rare, patients with congenital myashtenic syndrome may develop also autoimmune MG. The association of CMS with MG and thymoma is even much rarer to find. The diagnoses of the two conditions have very relevant treatment implications. doi:10.1016/j.jneuroim.2014.08.119
32 Eosinophilic CNS vasculitis can mimic demyelinating disease of the brain and spinal cord Raphael Schneider, Jenny Tsai, Daniel Selchen, David Munoz University of Toronto, St. Michael's Hospital, Toronto, Canada A previously healthy 61 year-old Filipino man developed progressive ascending sensory loss up to the T8 dermatome and paraparesis over a period of four weeks. Spinal cord magnetic resonance imaging (MRI) revealed a T2-hyperintense lesion from the T7 to the T10 spinal segment with varying degrees of gadolinium enhancement. Brain MRI imaging revealed several ovoid T2-hyperintense lesions in juxtacortical and periventricular areas. Cerebrospinal fluid analysis only showed a mild lymphocytic pleocytosis without atypical cells. Oligoclonal bands were not present. No microorganism was detected. He received five days of intravenous high-dose solu-medrol followed by five sessions of plasmaexchange (PLEX) for presumed central nervous system (CNS) demyelination, without a clinical response. Brain biopsy showed vasculitis involving small arteries with transmural infiltration by macrophages, lymphocytes, polymorphonuclear leukocytes, and numerous eosinophils destroying the vessel wall. Luxol fast blue stain revealed only minimal perivascular myelin loss. He was diagnosed with eosinophilic vasculitis and intravenous cyclophosphamide was started. Eosinophilic vasculitis is a feature of certain rheumatological conditions. Stroke caused by vasculitis is well described in the Churg Strauss syndrome, which is characterized by multisystem granulomatous eosinophilic vasculitis and peripheral eosinophilia. Our case illustrates that eosinophilic CNS vasculitis should be considered in the differential diagnosis of CNS demyelination, especially if highdose steroid treatment and PLEX fail to alter the clinical picture. doi:10.1016/j.jneuroim.2014.08.120
55 The comparison of mesenchymal stem cell with Mitoxantrone in treatment of patients with multiple sclerosis Saeed Shahbeigia, Mandana Mohyeddin Bonaba, Behrouz Nikbina, Yadollah Shakibaa, Ghazal Jalilzadehb, Abbas Najaric, Leila Cheginid, Torach Orange, Shaghayegh Ranjbarf, Samaneh Sheikhi Kouhsara, Foroogh Nejatollahig, Masoud Yousefvandh, Gholamreza Bakhshandepouri, Saeed Esmaeil Soofianj a
Immunology Department, Tehran, Iran; bJondishapour Neurology Clinic, Jondishapour Neurology Clinic, Tehran, Iran; cMinistry of Health, Ministry of Health, Tehran, Iran; dIran Medical University, Tehran, Iran; e Iran Medical University, Tehran, Iran; fShahid Beheshti Medical University, Shahid Beheshti Medical University, Tehran, Iran; gShiraz Medical University, Shiraz Medical University, Shiraz, Iran; hMinistry of
46
Abstracts
Health, Tehran, Iran; iPaytakht Mri Center, Paytakht Mri Center, Tehran, Iran; jIran Medical University, Tehran, Iran Introduction: Multiple sclerosis (MS) is an autoimmune-mediated disease of unknown etiology. There are two generally accepted options for treating MS. First, preventing the damage and second, to repair the already damaged in the Central Nervous System (CNS). Endogenous adult human stem cells can remyelinate the human CNS; however generally, this process often fails or is inhibited, resulting in chronic demyelination and progressive axonal death. In contrast, stem cells (SC) emerge as a potential new hope for treatment of MS. There is a growing body of literature that supports SC's potential contribution to immunomodulation and remyelination. We are going to examine the efficacy of MSC in treatment of the patients with MS and compare it with Mitoxantrone. Methods and materials: This is a double blind clinical trial study. In this study, 150 patients with MS were selected. The MS patients were aggressive type of MS who did not have any response to conventional DMDs. They have EDSS more than 4 and less than 7. These group of the patients were divided into 2 groups: 50 will be received autologous mesenchymal stem cells (A-MSC) the second group will be infused Mitoxantrone. In the first group, after harvesting of the bone marrow, we have purified MSC and cultured them. The first injection of cells was done after the MSC number reached to 30 million cells. In fact, we are going to inject 4 times MSCs. Except for the first dose; all injections will be done from L4 to L5 space intra-techally. For the other arm, Mitoxantrone have been infused in 0, 1 and 2 months and then every 3 months up to 140 mg/body m3. Clinical assessments, radiological findings and immunological markers will be evaluated before and after MSC and Mitoxantrone infusions as double blind evaluation. Results: In the paper, we will discuss the preliminary results of clinical, radiological and immunological assessments of the patients.
several new MS-treatments impact on the NK status, emphasising the need to investigate their role in the pathophysiology of MS. Material and Methods: Participants included untreated relapsing– remitting (RRMS) patients and healthy controls (HC). PBMCs were collected and the frequency of both NK-subsets analysed. As NK cells have cytotoxic and pro-inflammatory properties, the aim of this study was to stimulate both NK-subsets with the erythrocyte tumour cell line (K562 cells) and to assess their cytotoxicity by measuring CD107 expression and to stimulate both NK-subsets with IL-12 and IL-18 cytokines to measure their gamma interferon production. Results: We found unaltered frequencies of CD56+CD3− NK cells in RRMS patients. Furthermore, the functional activity (interferon-γ production, degranulation) of both NK subsets did not differ in RRMS compared to healthy controls. Conclusion: Our study concludes that there are no significant changes in NK cell frequency and functionality of the CD56bright and CD56dim NK subsets in untreated RRMS patients and we are awaiting NK cell data from our Alemtuzumab cohort to assess its impact on this population.
doi:10.1016/j.jneuroim.2014.08.122
24 Antiphospholipid antibodies in patients with multiple sclerosis or neuromyelitis optica Kazuya Takahashi, Kiyonobu Komai, Chiho Ishida, Yuko Motozaki, Tokuhei Ikeda, Ichiro Nozaki Department of Neurology, National Hospital Organization Iou Hospital, Kanazawa, Japan
doi:10.1016/j.jneuroim.2014.08.121
252 Unaltered frequency and functionality of CD56bright and CD56dim natural killer cells in untreated relapsing–remitting multiple sclerosis patients Sofia Sisaya, Alicia Roselloa, Korinna Henseleita, Matt Tana, Monica Marta-caldoa, Priya Duaa, Gary Warnesa, Jacqueline Palaceb, Gavin Giovannonia, Ute-christiane Meiera a
Centre of Neuroscience and Trauma, Blizard Building, Barts and The London School of Medicine and Dentistry, Queen Mary University, London, United Kingdom; bThe Department of Clinical Neurology, Oxford University Hospitals NHS Trust, Oxford, Oxford University Hospital, Oxford, United Kingdom Background: Multiple sclerosis (MS) is the most common disabling neurological disease in young adults in the western world. It is an inflammatory and neurodegenerative disorder of the central nervous system. The innate immune response in MS remains relatively poorly characterised and natural killer (NK) cells play a pivotal role. They contribute to innate host defence by cytolysis and production of cytokines such as gamma interferon. CD56+CD3− NK cells constitute ~ 13% of peripheral blood mononuclear cells (PBMCs) and can be divided into two subpopulations according to the expression level of CD56. Most NK-cells (90%) are CD56dim and the main mediators of cytotoxicity. A minority (10%) are CD56bright and an important source of immunoregulatory cytokines. Recent data implicated, that
Background: Antiphospholipid syndrome (APS) is characterized by arterial and venous thrombosis and the presence of antiphospholipid antibodies (aPLs), including anticardiolipin antibody (aCL) and lupus anticoagulant (LAC). However, some patients with multiple sclerosis (MS) continuously test positive for aPLs. Objective: We investigated the frequency of aPL positivity in patients with both MS and neuromyelitis optica (NMO) without APS-related clinical events, such as arterial and venous thrombosis. Subjects: Fifty patients fulfilled the 2010 inclusion criteria for MS in 2010 and 14 of those patients fulfilled the 2006 revised NMO criteria. Results: Eighteen percent of the patients with MS and 14% with NMO were continuously positive for 1 or more aPLs. Although phosphatidylserine/prothrombin complex antibodies (PS/PT) were found in both groups, LAC and beta2-glycoprotein I (GPI)-independent aCL were only positive in patients with MS. When LAC was positive in patients with MS, it was only demonstrated with the kaolin clotting time (KCT) assays, not the dilute Russell's viper venom time (dRVVT) assays. No patient in either group was positive for GPI-dependent aCL. Interestingly, 8% of patients with MS and 14% with NMO were temporarily positive for aCL or LAC. Discussion: The frequencies of aPL positivity were similar to those in previous reports; moreover, all of the patients exhibited low titers. Interestingly, although PS/PT is a key antibody for LAC, all patients with MS and LAC positivity tested negative for PS/PT. This result indicated that unknown autoreactive antibodies other than aPL may have cross-reactivity with LAC.
doi:10.1016/j.jneuroim.2014.08.123
sleep disorder was the initial symptom in four patients and in the rest it was preceded by gait instability. Patients developed variable degrees of chorea, dysautonomia, dysarthria, dysphagia, central hypoventilation. All four patients tested had HLA-DRB1*1001 and HLA-DQB1*0501 alleles. All patients' serum/CSF showed the same pattern of rat brain neuropil immunostaining that could be abrogated by immunocompetition assays suggesting a similar epitope, and they also labeled the cell membrane of live neurons. Patients' antibodies were IgG4. The immunoprecipitated target antigen was IgLON5, a neuronal adhesion molecule that is part of the immunoglobulin superfamily. The IgLON family of proteins is involved in the control of neuronal pathfinding and synaptogenesis. Patients had only IgLON5 antibodies and they did not have any additional antibodies against other neuronal surface antigens or IgLON family members. One of the 298 controls who had progressive supranuclear palsy had IgLON5 antibodies in serum but not in CSF. The neuropathological examination in two patients showed a neuronal tauopathy mainly involving the hypothalamus and tegmentum of the brainstem without inflammation. IgLON5 antibodies identify a novel sleep disorder characterized by NREM and REM parasomnia and stridor with sleep apnea with pathological findings suggesting a novel tauopathy. These findings may provide a link between autoimmunity and neurodegeneration.
doi:10.1016/j.jneuroim.2014.08.118
58 Myasthenia gravis associated with thymoma in a patient with CHRNE congenital myasthenic syndrome Ernestina Santosa, Isabel Moreirab, Maria Isabel Leitec, Jose Lopes Limab a Neurology Department, Hospital Santo Antonio, Instituto de Ciencias Biomedicas Abel Salazar, Universidade do Porto, Porto, Portugal; bNeurology Department, Hospital Santo Antonio, Porto, Portugal; cDepartment of Clinical Neurology, Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom
Background: The dysfunction or loss of acetylcholine receptors (AChR) in the neuromuscular junction may have different causes. More frequently the cause is autoimmune (myasthenia gravis, MG) — which is associated with antibodies to AChR. Less commonly the disease is genetic (congenital myasthenic syndromes; CMS). Case Report: Two brothers, with non-consanguineous healthy parents, have, since the age of 4 and 6 years, ophthalmoplegia, dysphonia, fatigable difficulty in chewing and fatigable weakness in the limb muscles. Electromyography showed significant decrement in the amplitude of CMAP with repetitive stimulation, suggestive of pos-synaptic defect of neuromuscular transmission. Muscle enzymes were normal. Muscle biopsy performed in one of the brothers showed subtle abnormalities suggestive of myopathic origin. Genetic study revealed a mutation on the gene CHRNE, confirming the diagnosis of congenital myasthenic syndrome in both children. They started treatment with pyridostigmine with improvement. One of the brothers was stable until the age of 34 (2003). At that time he developed dyspnoea and worsening of the weakness in the limbs. Chest CT scan showed a mediastinal mass and several adenopathies. Thymus was removed, and the histology revealed a thymoma. The adenopathies were found to be of a thyroid papillary carcinoma. He was submitted also to thyroidectomy followed by radiotherapy. Some months later he improved returning to his baseline. Antibodies anti-AChR's were not searched during the period of the worsening and previously to the thymectomy. In 2012 they were negative.
45
Comments: These cases demonstrate that, although very rare, patients with congenital myashtenic syndrome may develop also autoimmune MG. The association of CMS with MG and thymoma is even much rarer to find. The diagnoses of the two conditions have very relevant treatment implications. doi:10.1016/j.jneuroim.2014.08.119
32 Eosinophilic CNS vasculitis can mimic demyelinating disease of the brain and spinal cord Raphael Schneider, Jenny Tsai, Daniel Selchen, David Munoz University of Toronto, St. Michael's Hospital, Toronto, Canada A previously healthy 61 year-old Filipino man developed progressive ascending sensory loss up to the T8 dermatome and paraparesis over a period of four weeks. Spinal cord magnetic resonance imaging (MRI) revealed a T2-hyperintense lesion from the T7 to the T10 spinal segment with varying degrees of gadolinium enhancement. Brain MRI imaging revealed several ovoid T2-hyperintense lesions in juxtacortical and periventricular areas. Cerebrospinal fluid analysis only showed a mild lymphocytic pleocytosis without atypical cells. Oligoclonal bands were not present. No microorganism was detected. He received five days of intravenous high-dose solu-medrol followed by five sessions of plasmaexchange (PLEX) for presumed central nervous system (CNS) demyelination, without a clinical response. Brain biopsy showed vasculitis involving small arteries with transmural infiltration by macrophages, lymphocytes, polymorphonuclear leukocytes, and numerous eosinophils destroying the vessel wall. Luxol fast blue stain revealed only minimal perivascular myelin loss. He was diagnosed with eosinophilic vasculitis and intravenous cyclophosphamide was started. Eosinophilic vasculitis is a feature of certain rheumatological conditions. Stroke caused by vasculitis is well described in the Churg Strauss syndrome, which is characterized by multisystem granulomatous eosinophilic vasculitis and peripheral eosinophilia. Our case illustrates that eosinophilic CNS vasculitis should be considered in the differential diagnosis of CNS demyelination, especially if highdose steroid treatment and PLEX fail to alter the clinical picture. doi:10.1016/j.jneuroim.2014.08.120
55 The comparison of mesenchymal stem cell with Mitoxantrone in treatment of patients with multiple sclerosis Saeed Shahbeigia, Mandana Mohyeddin Bonaba, Behrouz Nikbina, Yadollah Shakibaa, Ghazal Jalilzadehb, Abbas Najaric, Leila Cheginid, Torach Orange, Shaghayegh Ranjbarf, Samaneh Sheikhi Kouhsara, Foroogh Nejatollahig, Masoud Yousefvandh, Gholamreza Bakhshandepouri, Saeed Esmaeil Soofianj a
Immunology Department, Tehran, Iran; bJondishapour Neurology Clinic, Jondishapour Neurology Clinic, Tehran, Iran; cMinistry of Health, Ministry of Health, Tehran, Iran; dIran Medical University, Tehran, Iran; e Iran Medical University, Tehran, Iran; fShahid Beheshti Medical University, Shahid Beheshti Medical University, Tehran, Iran; gShiraz Medical University, Shiraz Medical University, Shiraz, Iran; hMinistry of
46
Abstracts
Health, Tehran, Iran; iPaytakht Mri Center, Paytakht Mri Center, Tehran, Iran; jIran Medical University, Tehran, Iran Introduction: Multiple sclerosis (MS) is an autoimmune-mediated disease of unknown etiology. There are two generally accepted options for treating MS. First, preventing the damage and second, to repair the already damaged in the Central Nervous System (CNS). Endogenous adult human stem cells can remyelinate the human CNS; however generally, this process often fails or is inhibited, resulting in chronic demyelination and progressive axonal death. In contrast, stem cells (SC) emerge as a potential new hope for treatment of MS. There is a growing body of literature that supports SC's potential contribution to immunomodulation and remyelination. We are going to examine the efficacy of MSC in treatment of the patients with MS and compare it with Mitoxantrone. Methods and materials: This is a double blind clinical trial study. In this study, 150 patients with MS were selected. The MS patients were aggressive type of MS who did not have any response to conventional DMDs. They have EDSS more than 4 and less than 7. These group of the patients were divided into 2 groups: 50 will be received autologous mesenchymal stem cells (A-MSC) the second group will be infused Mitoxantrone. In the first group, after harvesting of the bone marrow, we have purified MSC and cultured them. The first injection of cells was done after the MSC number reached to 30 million cells. In fact, we are going to inject 4 times MSCs. Except for the first dose; all injections will be done from L4 to L5 space intra-techally. For the other arm, Mitoxantrone have been infused in 0, 1 and 2 months and then every 3 months up to 140 mg/body m3. Clinical assessments, radiological findings and immunological markers will be evaluated before and after MSC and Mitoxantrone infusions as double blind evaluation. Results: In the paper, we will discuss the preliminary results of clinical, radiological and immunological assessments of the patients.
several new MS-treatments impact on the NK status, emphasising the need to investigate their role in the pathophysiology of MS. Material and Methods: Participants included untreated relapsing– remitting (RRMS) patients and healthy controls (HC). PBMCs were collected and the frequency of both NK-subsets analysed. As NK cells have cytotoxic and pro-inflammatory properties, the aim of this study was to stimulate both NK-subsets with the erythrocyte tumour cell line (K562 cells) and to assess their cytotoxicity by measuring CD107 expression and to stimulate both NK-subsets with IL-12 and IL-18 cytokines to measure their gamma interferon production. Results: We found unaltered frequencies of CD56+CD3− NK cells in RRMS patients. Furthermore, the functional activity (interferon-γ production, degranulation) of both NK subsets did not differ in RRMS compared to healthy controls. Conclusion: Our study concludes that there are no significant changes in NK cell frequency and functionality of the CD56bright and CD56dim NK subsets in untreated RRMS patients and we are awaiting NK cell data from our Alemtuzumab cohort to assess its impact on this population.
doi:10.1016/j.jneuroim.2014.08.122
24 Antiphospholipid antibodies in patients with multiple sclerosis or neuromyelitis optica Kazuya Takahashi, Kiyonobu Komai, Chiho Ishida, Yuko Motozaki, Tokuhei Ikeda, Ichiro Nozaki Department of Neurology, National Hospital Organization Iou Hospital, Kanazawa, Japan
doi:10.1016/j.jneuroim.2014.08.121
252 Unaltered frequency and functionality of CD56bright and CD56dim natural killer cells in untreated relapsing–remitting multiple sclerosis patients Sofia Sisaya, Alicia Roselloa, Korinna Henseleita, Matt Tana, Monica Marta-caldoa, Priya Duaa, Gary Warnesa, Jacqueline Palaceb, Gavin Giovannonia, Ute-christiane Meiera a
Centre of Neuroscience and Trauma, Blizard Building, Barts and The London School of Medicine and Dentistry, Queen Mary University, London, United Kingdom; bThe Department of Clinical Neurology, Oxford University Hospitals NHS Trust, Oxford, Oxford University Hospital, Oxford, United Kingdom Background: Multiple sclerosis (MS) is the most common disabling neurological disease in young adults in the western world. It is an inflammatory and neurodegenerative disorder of the central nervous system. The innate immune response in MS remains relatively poorly characterised and natural killer (NK) cells play a pivotal role. They contribute to innate host defence by cytolysis and production of cytokines such as gamma interferon. CD56+CD3− NK cells constitute ~ 13% of peripheral blood mononuclear cells (PBMCs) and can be divided into two subpopulations according to the expression level of CD56. Most NK-cells (90%) are CD56dim and the main mediators of cytotoxicity. A minority (10%) are CD56bright and an important source of immunoregulatory cytokines. Recent data implicated, that
Background: Antiphospholipid syndrome (APS) is characterized by arterial and venous thrombosis and the presence of antiphospholipid antibodies (aPLs), including anticardiolipin antibody (aCL) and lupus anticoagulant (LAC). However, some patients with multiple sclerosis (MS) continuously test positive for aPLs. Objective: We investigated the frequency of aPL positivity in patients with both MS and neuromyelitis optica (NMO) without APS-related clinical events, such as arterial and venous thrombosis. Subjects: Fifty patients fulfilled the 2010 inclusion criteria for MS in 2010 and 14 of those patients fulfilled the 2006 revised NMO criteria. Results: Eighteen percent of the patients with MS and 14% with NMO were continuously positive for 1 or more aPLs. Although phosphatidylserine/prothrombin complex antibodies (PS/PT) were found in both groups, LAC and beta2-glycoprotein I (GPI)-independent aCL were only positive in patients with MS. When LAC was positive in patients with MS, it was only demonstrated with the kaolin clotting time (KCT) assays, not the dilute Russell's viper venom time (dRVVT) assays. No patient in either group was positive for GPI-dependent aCL. Interestingly, 8% of patients with MS and 14% with NMO were temporarily positive for aCL or LAC. Discussion: The frequencies of aPL positivity were similar to those in previous reports; moreover, all of the patients exhibited low titers. Interestingly, although PS/PT is a key antibody for LAC, all patients with MS and LAC positivity tested negative for PS/PT. This result indicated that unknown autoreactive antibodies other than aPL may have cross-reactivity with LAC.
doi:10.1016/j.jneuroim.2014.08.123