A modified application of the luciferase immunoprecipitation systems for detecting antibodies to the G protein-coupled receptors

Background:　When multipass transmembrane molecules are located on the cell surface, there may be interaction with notonly bioactive molecules but also pathogenic molecules in areas protruding outside the cell. In antibody-mediated autoimmunedisorders, it has been found that the autoantibodies occasionally attack membrane molecules on the cell surface, thus causingthe disease such as myasthenia gravis. In such cases, highly sensitive autoantibody detection technology is required for earlydiagnosis. However, autoantibody analysis technology that is specialized for membrane molecules is still under development.Here we demonstrate a novel method for detecting of antibodies against the extracellular portions of multipass transmembranemolecules.Methods:　Antibodies for muscarinic acetylcholine receptor type3 (M3R) were detected with two kinds of luciferase immunoprecipitaionsystems (LIPS), conventional LIPS (cLIPS) and its modified application, termed modified LIPS (mLIPS). In mLIPS, antibodiesagainst extracellular portions of membrane molecules could be preferentially detected.Results:　An antibody to the amino-terminal portion of human M3R was detected with modified LIPS with a high sensitivity. Incontrast, an antibody to the carboxyl-terminal portion was not detected with mLIPS, because it did not interact with intracellularportions of M3R in living cells. We also found antibodies for M3R in a patient serum with Sjogren’s syndrome.Conclusion:　Our technology has a promising future, and we hope that it will be applied in the analysis of antibodies against adiverse range of multipass transmembrane molecules, including GPCRs

Gait and posture assessments of a patient treated with deep brain stimulation in dystonia using three-dimensional motion analysis systems

Kinesiologic analysis of gait disorders, postural instabilities and abnormal movements is quite difficult to assess objectively by clinical observation, such as by specific scale and video recordings. In this study, we reported one of the aspects of the usefulness of three-dimensional motion analysis (Vicon Systems, Oxford, United Kingdom), which can measure inclusive data of movement disorders and substitute for conventional assessments. A 49-year-old man who had various dystonic symptoms, mainly on his left side of the body, responded well to deep brain stimulation (DBS). The examination quantified how the involuntary movements or other symptoms with dystonia changed before and after treatments

高齢で発症した重症筋無力症は重症化しやすい

Objective: The continuous increase in the number of patients presenting with late-onset myasthenia gravis (LOMG) underscores the need for a better understanding of the clinical course and the establishment of an optimal therapeutic strategy. We aimed to clarify factors associated with clinical outcomes in LOMG. Methods: We retrospectively reviewed the clinical profiles of 40 patients with early-onset MG (EOMG) (onset age: 49 years or younger), 30 patients with non-elderly LOMG (onset age: 50–64 years), and 28 patients with elderly LOMG(onset age: 65 years or older) and compared the subgroups according to onset age and thymus status. The evaluated parameters were MGFA classification before treatment, MG-ADL score, complicating diseases, antibody titer, treatment, and MGFA post-intervention status. Results: Elderly LOMG patients showed transition to generalized symptoms at a higher frequency and underwent thymectomyless frequently than EOMG and non-elderly LOMG patients (p < 0.001). The frequencies of crisis and plasmapheresis were significantly lower in thymectomized LOMG patients without thymoma than in thymectomized LOMG patients with thymoma or non-thymectomized LOMG patients (p < 0.01, P < 0.05, respectively). However, the outcome was not significantly different. All of the thymectomized LOMG patients without thymoma presenting with hyperplasia or thymic cyst had a favorable clinical course. Conclusions: Our study showed that elderly LOMG patients are more prone to severity, suggesting that they require aggressive immunomodulatory therapy

Clinical Features of Autoimmune Autonomic Ganglionopathy and the Detection of Subunit-Specific Autoantibodies to the Ganglionic Acetylcholine Receptor in Japanese Patients

Autoimmune autonomic ganglionopathy (AAG) is a rare acquired channelopathy that is characterized by pandysautonomia, in which autoantibodies to ganglionic nicotinic acetylcholine receptors (gAChR) may play a central role. Radioimmunoprecipitation (RIP) assays have been used for the sensitive detection of autoantibodies to gAChR in the serum of patients with AAG. Here, we developed luciferase immunoprecipitation systems (LIPS) to diagnose AAG based on IgGs to both the α3 and β4 gAChR subunits in patient serum. We reviewed the serological and clinical data of 50 Japanese patients who were diagnosed with AAG. With the LIPS testing, we detected anti-α3 and -β4 gAChR antibodies in 48% (24/50) of the patients. A gradual mode of onset was more common in the seropositive group than in the seronegative group. Patients with AAG frequently have orthostatic hypotension and upper and lower gastrointestinal tract symptoms, with or without anti-gAChR. The occurrence of autonomic symptoms was not significantly different between the seropositive and seronegative group, with the exception of achalasia in three patients from the seropositive group. In addition, we found a significant overrepresentation of autoimmune diseases in the seropositive group and endocrinological abnormalities as an occasional complication of AAG. Our results demonstrated that the LIPS assay was a useful novel tool for detecting autoantibodies against gAChR in patients with AAG

Current practices and factors affecting evidence-based occupational therapy practice for patients with dementia in Japan

Objective/background The rapidly increasing incidence of patients with dementia in Japan is creating an urgent demand for evidence-based occupational therapy (EBOT), which has been reported to improve clinical efficacy. This study aimed to examine the current practice of EBOT for patients with dementia in Japan and clarify factors influencing its application. Methods We conducted an anonymous, self-administered questionnaire survey by mail. The participants were 432 occupational therapists treating patients with dementia at 432 designated medical institutions nationwide. Descriptive statistics were calculated, and multiple regression analysis was performed to clarify the factors influencing the present application of EBOT. Results The response rate was 31.3%. Among the participants, 46.3% responded 'somewhat frequently' or more (5, 6, and 7 on a 7-grade scale) to a question on the frequency of practice of EBOT. Using multiple regression analysis with the stepwise method, we selected the model with the highest degree of fit. This model extracted three factors, namely, ability to understand scientific papers, sufficiency of means of getting information, and availability of advice, which had standardized partial regression coefficients (beta) of 0.419, 0.214, and 0.158, respectively. Conclusions The three factors extracted using the multiple regression analysis indicate that in order to encourage EBOT for patients with dementia, occupational therapists need to acquire reading comprehension skills, so that they can assess the quality of scientific papers. Furthermore, it is important to create environments where they have access to publications and can discuss research with superiors and colleagues

MRS of HDLS and CSF1R Mutation Carrier

Purpose: Hereditary diffuse leukoencephalopathy with spheroids (HDLS) is a rare neurodegenerative disorder with various clinical presentations. Mutation of the colony-stimulating factor 1 receptor (CSF1R) gene is considered to be a cause of this autosomal dominant disorder. The purpose of this study was to report magnetic resonance spectroscopy (MRS) findings in patients with HDLS and asymptomatic carriers and to clarify the use of MRS in this disease. Materials and Methods: In this retrospective, institutional review board-approved study, we included four consecutive patients, genetically diagnosed with HDLS, and two asymptomatic carriers after acquiring informed consent. We performed single-voxel MRS of the left centrum semiovale on a 3T clinical scanner. We also included a sex-matched normal dataset. We quantified N-acetylaspartate (NAA), creatine, choline-containing compounds (Cho), glutamine, glutamate (Glu), myo-inositol (Ins), glutathione, lactate (Lac), and gamma-amino butyric acid using LCModel. We performed statistical analysis, and P value <0.05 was considered significant. Results: In HDLS cases, MRS revealed decreased NAA and Glu concentrations, which probably reflected neuronal damage and/or loss, and a subsequent reduction of neurotransmitters. A patient with HDLS also had increased Cho and Ins concentrations, indicating gliosis, and increased Cho concentration was also observed in an asymptomatic carrier. This suggests that metabolic changes had already occurred in an asymptomatic state. Conclusion: We demonstrated changes in metabolite concentrations not only in patients with HDLS but also in asymptomatic CSF1R mutation carriers. Our study indicates that MRS is a potentially useful tool for the analysis of metabolic and pathophysiological findings of HDLS, even during the early stages of disease