RESUMEN
Heat-shock proteins (HSPs) have been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). In this study, we aimed to examine whether the serum levels of HSPs (HSP27, HSP70, and HSP90) are altered in patients with ALS. We included 58 patients diagnosed with ALS and 85 control individuals. Serum HSP levels of patients and controls were determined using enzyme-linked immunosorbent assay. The serum levels of HSP70 and HSP90 were significantly higher in patients than in controls. In contrast, serum levels of HSP27 did not differ significantly between the patient and control groups. Moreover, serum levels of HSP70 and HSP90 in patients remained high throughout the duration of the disease. Taken together, our findings suggest that HSPs might have a role in ALS progression throughout the course of the disease. Further studies are needed to clarify the role of HSPs in the pathogenesis of ALS.
Asunto(s)
Esclerosis Amiotrófica Lateral/sangre , Proteínas de Choque Térmico/sangre , Anciano , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Femenino , Proteínas de Choque Térmico HSP27 , Proteínas HSP70 de Choque Térmico , Proteínas HSP90 de Choque Térmico , Humanos , Masculino , Persona de Mediana EdadRESUMEN
While G-protein-coupled receptors (GPCRs) represent the largest class of cell surface proteins, there are ≥100 orphan GPCRs whose endogenous ligands are unknown. Accordingly, these could prove to be potential therapeutic targets for the pharmaceutical intervention of various diseases. Constitutively active orphan GPCRs are activated without ligands; thus, inverse agonists may be very useful pharmacological tools for inhibiting constitutive activity. However, in general, inverse agonist screening is considered more difficult to perform with high quality than antagonist screening, particularly due to the narrow assay window. We developed a high-throughput screening (HTS)-compatible assay to identify inverse agonists of GPR3. GPR3 is expressed in the central nervous system (CNS) and is known to be related to Alzheimer's disease and other CNS diseases. The GPR3 inducible cell line was established using T-REx 293 cells that stably expressed the tetracycline repressor protein, and the cAMP biosensor, GloSensor, was stably co-expressed. After optimization of the induction level of GPR3 and assay conditions, the GloSensor assay showed an approximately 20-fold signal-to-background ratio and high sensitivity. Using the HTS method, we successfully screened a library of hundreds of thousands of compounds for the inhibition of constitutive activity with good quality and excellent reproducibility. Finally, 35 compounds were identified as GPR3 selective inverse agonists. This inverse agonist screening approach using GloSensor in combination with the inducible expression of orphan GPCR indicates universal applicability to the search for inverse agonists of constitutively active orphan GPCRs.
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Técnicas Biosensibles , Agonismo Inverso de Drogas , Ensayos Analíticos de Alto Rendimiento , Receptores Acoplados a Proteínas G/agonistas , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/genética , AMP Cíclico/genética , Humanos , Ligandos , Unión Proteica/efectos de los fármacos , Receptores Acoplados a Proteínas G/genéticaRESUMEN
O,O,S-Trimethylphosphorothioate (OOS-TMP) has been shown to induce hypophagia and hypopraxia. Recent studies suggest that OOS-TMP-induced anorexia is partly mediated by its effect on the central nervous system. In this study, we examined the profiles of N-acylethanolamines (NEAs), including five amide-linked compounds, in the gastrointestinal system in C57BL/6J (B6) mice. The present results shown an orexigenic profile of the levels of NEAs with downregulation of the anorectic lipid, N-stearoylethanolamine (SEA), upregulation of the orexigenic lipid, 2-arachidonoyl glycerol (2-AG), at 2 h and upregulation of 2-AG at 24 h albeit with significant anorexia. However, the data indicated that the high level of 2-AG may be responsible for the hypopraxia. We next explored whether OOS-TMP may affect two models of hyperphagia and hyperglycemia, ins2(+/Akita) B6 (Akita) and B6-lepr(db)/lepr(db) mice (db/db). We identified potential anorexigenic effects in B6, Akita and db/db mice. Moreover, OOS-TMP was found to reduce blood glucose in Akita mice but not in db/db mice. Collectively, these findings suggest that N-acylethanolamines are not involved in the hypophagia but rather hypopraxia, and may play multiple physiological roles in this process. OOS-TMP might be a promising candidate for anti-obesity and anti-diabetic drug development.
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Fármacos Antiobesidad , Etanolaminas/metabolismo , Hiperglucemia/tratamiento farmacológico , Hiperfagia/tratamiento farmacológico , Hipoglucemiantes , Insecticidas/farmacología , Obesidad/tratamiento farmacológico , Organotiofosfatos/farmacología , Animales , Depresores del Apetito/farmacología , Glucemia/metabolismo , Ingestión de Alimentos/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Hiperfagia/psicología , Inyecciones Intraventriculares , Insecticidas/administración & dosificación , Mucosa Intestinal/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Obesidad/etiología , Obesidad/psicología , Organotiofosfatos/administración & dosificación , Aumento de Peso/efectos de los fármacosRESUMEN
In Japan, a significant number of adolescent girls complained unusual symptoms after human papillomavirus (HPV) vaccination, and the vast majority of them were initially diagnosed as having psychiatric illness because of the absence of pathologic findings, radiological images and specific abnormalities in laboratory test results. Later, these symptoms were supposed to be adverse effects after HPV vaccination, and the recommendation for HPV vaccination was withdrawn by Japanese Ministry of Public Health, Labour and Welfare 4 years and 9 months ago. However, a causal link has not been demonstrated between HPV vaccination and the development of these symptoms. Our study has shown that the period of HPV vaccination considerably overlapped with that of unique postvaccination symptom development, adding that new patients with possible HPV vaccine-related symptoms have not appeared during our recent 28-month follow-up period. This social episode has now subsided in Japan.
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Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/etiología , Papillomaviridae/inmunología , Vacunas contra Papillomavirus/efectos adversos , Vacunación/efectos adversos , Sistemas de Registro de Reacción Adversa a Medicamentos , Humanos , JapónRESUMEN
The nuclear factor κB (NF-κB) pathway is critical for regulating immune and inflammatory responses, and uncontrolled NF-κB activation is closely associated with various inflammatory diseases and malignant tumors. The Met1-linked linear ubiquitin chain, which is generated by linear ubiquitin chain assembly complex (LUBAC), is important for regulating NF-κB activation. This process occurs through the linear ubiquitination of NF-κB essential modulator, a regulatory subunit of the canonical inhibitor of the NF-κB kinase complex. In this study, we have established a robust and efficient high-throughput screening (HTS) platform to explore LUBAC inhibitors, which may be used as tool compounds to elucidate the pathophysiological role of LUBAC. The HTS platform consisted of both cell-free and cell-based assays: (1) cell-free LUBAC-mediated linear ubiquitination assay using homogenous time-resolved fluorescence technology and (2) cell-based LUBAC assay using the NF-κB luciferase reporter gene assay. By using the HTS platform, we performed a high-throughput chemical library screen and identified several hit compounds with selectivity against a counterassay. Liquid chromatography-mass spectrometry analysis revealed that these compounds contain a chemically reactive lactone structure, which is transformed to give reactive α,ß-unsaturated carbonyl compounds. Further investigation revealed that the reactive group of these compounds is essential for the inhibition of LUBAC activity.
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Descubrimiento de Drogas , Fluoroinmunoensayo/métodos , Ensayos Analíticos de Alto Rendimiento , Ubiquitina/metabolismo , Ubiquitinación/efectos de los fármacos , Línea Celular , Sistema Libre de Células , Cromatografía Liquida , Descubrimiento de Drogas/métodos , Expresión Génica , Genes Reporteros , Humanos , Espectrometría de Masas , FN-kappa B/metabolismoRESUMEN
The secondary process of lipid peroxidation proceeds by the free radical reaction to produce some toxic aldehydes. Since γ-tocopherol (γ-TH), one of the major forms of vitamin E in some vegetable oils, acts as a free radical scavenger, γ-TH may suppress the formation of such aldehydes. This study reports the effect and reaction products of γ-TH on the hemin- or myoglobin-catalyzed decomposition of 1-palmitoyl-2-linoleoyl-3-sn-phosphatidylcholine 13-hydroperoxide (PLPC-OOH) in micelles and liposomes. γ-TH and PLPC-OOH in micelles were reacted in the presence of hemin, and the reaction products were characterized as 1-palmitoyl-2-[(8a-dioxy-γ-tocopherone)-12,13-epoxyoctadecenoyl]-3-sn-phosphatidylcholines (γT-OO-epoxyPLPC), 1-palmitoyl-2-[(γ-tocopheroxy)-12,13-epoxyoctadecenoyl]-3-sn-phosphatidylcholines (γT-epoxyPLPC), and the adducts of γ-TH dimer with PLPC-OOH derived epoxyperoxyl and epoxyalkyl radicals (γTD-OO-epoxyPLPC and γTD-epoxyPLPC). The hemin- and myoglobin-catalyzed decomposition of PLPC-OOH in micelles produced hexanal and 4-hydroxy-2-nonenal as the major aldehydic products. γ-TH suppressed the formation of these aldehydes as the same level as α-TH did, and γ-tocopherylquinone, tocored, γ-TH dimers, and the addition products (γT-OO-epoxyPLPC, γT-epoxyPLPC, γTD-OO-epoxyPLPC, and γTD-epoxyPLPC) were formed during the reaction. In liposomes, hexanal was detected as the major aldehyde and its suppression by γ-TH was less effective than that by α-TH. The results indicate that γ-TH may suppress the formation of aldehydes by trapping the epoxyperoxyl and epoxyalkyl radicals derived from PLPC-OOH although its ability is weak in liposomal systems.
Asunto(s)
Hemina/química , Mioglobina/química , Fosfatidilcolinas/química , gamma-Tocoferol/química , Catálisis , Liposomas/química , MicelasRESUMEN
INTRODUCTION: In Japan, after receiving human papillomavirus vaccination, a significant number of adolescent girls experienced various symptoms, the vast majority of which have been ascribed to chronic regional pain syndrome, orthostatic intolerance, and/or cognitive dysfunction. However, a causal link has not been established between human papillomavirus vaccination and the development of these symptoms. OBJECTIVE: The aim of this study was to clarify the temporal relationship between human papillomavirus vaccination and the appearance of post-vaccination symptoms. METHODS: Between June 2013 and December 2016, we examined symptoms and objective findings in 163 female patients who had received human papillomavirus vaccination. We used newly defined diagnostic criteria for accurate inclusion of patients who experienced adverse symptoms after human papillomavirus vaccination; these diagnostic criteria were created for this study, and thus their validity and reliability have not been established. RESULTS: Overall, 43 female patients were excluded. Among the remaining 120 patients, 30 were diagnosed as having definite vaccine-related symptoms, and 42 were diagnosed as probable. Among these 72 patients, the age at initial vaccination ranged from 11 to 19 years (average 13.6 ± 1.6 years), and the age at appearance of symptoms ranged from 12 to 20 years (average 14.4 ± 1.7 years). The patients received the initial human papillomavirus vaccine injection between May 2010 and April 2013. The first affected girl developed symptoms in October 2010, and the last two affected girls developed symptoms in October 2015. The time to onset after the first vaccine dose ranged from 1 to 1532 days (average 319.7 ± 349.3 days). CONCLUSIONS: The period of human papillomavirus vaccination considerably overlapped with that of unique post-vaccination symptom development. Based on these sequential events, it is suggested that human papillomavirus vaccination is related to the transiently high prevalence of the previously mentioned symptoms including chronic regional pain syndrome and autonomic and cognitive dysfunctions in the vaccinated patients.
Asunto(s)
Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/epidemiología , Vacunas contra Papillomavirus/efectos adversos , Adolescente , Sistemas de Registro de Reacción Adversa a Medicamentos , Niño , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/etiología , Femenino , Humanos , Japón/epidemiología , Infecciones por Papillomavirus/prevención & control , Prevalencia , Análisis Espacio-Temporal , Neoplasias del Cuello Uterino/prevención & control , Vacunación/efectos adversos , Adulto JovenRESUMEN
Mycobacterium abscessus infection tends to occur in patients with an advanced immunocompromised status. We encountered a case of intractable cutaneous M. abscessus infection that developed in a patient with systemic lupus erythematosus (SLE) during maintenance therapy. A 28-year-old woman developed a fever and redness of the skin on her buttocks. General antibacterial therapy was ineffective, and acid-fast bacteria were detected in the biopsy that was conducted to differentiate the dermal symptoms of SLE. The clinical findings eventually improved; however, the symptoms recurred multiple times during treatment. Despite recent advances in SLE treatment, M. abscessus infection remains a considerable complication of SLE.
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Antibacterianos/uso terapéutico , Claritromicina/uso terapéutico , Lupus Eritematoso Sistémico/complicaciones , Lupus Eritematoso Sistémico/tratamiento farmacológico , Infecciones por Mycobacterium no Tuberculosas/tratamiento farmacológico , Infecciones por Mycobacterium no Tuberculosas/etiología , Micobacterias no Tuberculosas/efectos de los fármacos , Adulto , Femenino , Humanos , Huésped Inmunocomprometido/efectos de los fármacos , Infecciones por Mycobacterium no Tuberculosas/microbiología , Resultado del TratamientoRESUMEN
CONTEXT: Cerebrotendinous xanthomatosis (CTX) is an autosomal recessively inherited lipid storage disease caused by mutation in the CYP27A1 gene. Spinal form CTX is a rare clinical subgroup of CTX and only 14 patients from 11 families have been reported to date. Here, we report the first Asian patient with spinal form CTX showing characteristic radiological findings. FINDINGS: The patient, a 46-year-old Japanese male, developed sensory disturbance of the lower legs at 39 and spastic gait at 46 years of age. Spinal cord magnetic resonance imaging (MRI) revealed a long hyperintense lesion involving lateral corticospinal tracts and gracile tracts in the cervical and thoracic cord on T2-weighted images. Gallium-67 (67Ga) scintigraphy revealed abnormal uptake in the Achilles tendons and the serum cholestanol level was elevated. CYP27A1 gene analysis identified homozygous missense mutation, c.1214G>A (p.R405Q). The patient was treated with atorvastatin monotherapy, which reduced serum cholestanol to less than 50% of the pretreatment level. CONCLUSION: Spinal form CTX should be considered in the differential diagnosis of cryptogenic myelopathy, especially in patients with a long spinal cord lesion, as treatment with chenodeoxycholic acid and/or competitive inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase reverse the metabolic derangement and prevent the neurologiccal dysfunction.
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Tractos Piramidales/diagnóstico por imagen , Enfermedades de la Médula Espinal/diagnóstico , Xantomatosis Cerebrotendinosa/diagnóstico , Colestanotriol 26-Monooxigenasa/genética , Diagnóstico Diferencial , Humanos , Masculino , Persona de Mediana Edad , Mutación Missense , Tractos Piramidales/patología , Xantomatosis Cerebrotendinosa/tratamiento farmacológico , Xantomatosis Cerebrotendinosa/genéticaRESUMEN
Neurological involvement in relapsing polychondritis (RP) is relatively rare. We describe the case of an 80-year-old man who presented with hypertrophic pachymeningitis (HP) together with arthritis as the first manifestation of RP. Auricular chondritis, which subsequently determined the diagnosis of RP, occurred a few weeks after the detection of HP. The neurological symptoms, as well as arthritis, were promptly improved by treatment with corticosteroids. It is generally difficult to diagnose RP in the absence of typical cartilaginous involvement; however, the present case suggests that HP may occur as an early clinical manifestation of RP.
RESUMEN
The structure and the stability of various cyclopentadienides, which involve 6pi, 10pi, 14pi, and 22pi electrons, are investigated from computations at various levels of theory as well as from orbital interaction analyses. The reason that some of the cyclopentadienides are stabilized and others are destabilized by the introduction of aromatic rings is discussed in terms of absolute hardness and orbital interaction. Cyclopentadienide, a special 6pi-electron system, has the largest value of absolute hardness among the condensed cyclopentadienides investigated; thus this carbanion resists both oxidation and reduction most strongly. The absolute hardness decreases when aromatic rings are introduced to cyclopentadienide to form condensed cyclopentadienides, depending on the way they are connected. Computed values of the ionization potential and oxidation potentials measured in solution have a linear correlation within isomers of the same size, but are not in agreement for different sets of isomers. Solvent effects on the ionization potential are assessed by performing self-consistent reaction field calculations, the results being in excellent agreement with experiments. It is demonstrated that the solvent effects are significant in small cyclopentadienides of 6pi- and 10pi-electron systems, compared to larger ones and that addition of condensed aromatic rings intrinsically stabilizes the formed condensed cyclopentadienides with respect to ionization potential.
RESUMEN
Familial amyotrophic lateral sclerosis accounts for about 5% of all cases of the neurodegenerative disorder amyotrophic lateral sclerosis. Genetic mutations in Cu/Zn superoxide dismutase (SOD1) have been associated with one kind of familial amyotrophic lateral sclerosis (ALS1). We identified a novel duplication mutation in exon 1 of the SOD1 gene in a Japanese family whose members had lower motor neuron diseases. The patients showed slow disease progression, with the onset of lower limb muscle weakness and exertional dyspnea. Some patients had mild motor and sensory neuropathy and/or bladder dysfunction, which is further evidence that SOD1 mutation results in a predominantly lower motor neuron phenotype.
Asunto(s)
Exones/genética , Duplicación de Gen/genética , Estudios de Asociación Genética , Enfermedad de la Neurona Motora/genética , Mutación/genética , Superóxido Dismutasa/genética , Anciano , Pueblo Asiatico/genética , Progresión de la Enfermedad , Femenino , Humanos , Persona de Mediana Edad , Superóxido Dismutasa-1 , Factores de TiempoRESUMEN
OBJECTIVE: To investigate the causes of neurological manifestations in girls immunized with the human papillomavirus (HPV) vaccine. METHODS: During the past nine months, 44 girls visited us complaining of several symptoms after HPV vaccination. Four patients with other proven disorders were excluded, and the remaining forty subjects were enrolled in this study. RESULTS: The age at initial vaccination ranged from 11 to 17 years, and the average incubation period after the first dose of the vaccine was 5.47±5.00 months. Frequent manifestations included headaches, general fatigue, coldness of the legs, limb pain and weakness. The skin temperature examined in 28 girls with limb symptoms exhibited a slight decrease in the fingers (30.4±2.6 °C) and a moderate decrease in the toes (27.1±3.7 °C). Digital plethysmograms revealed a reduced height of the waves, especially in the toes. The limb symptoms of four girls were compatible with the Japanese clinical diagnostic criteria for complex regional pain syndrome (CRPS), while those in the other 14 girls were consistent with foreign diagnostic criteria for CRPS. The Schellong test identified eight patients with orthostatic hypotension and four patients with postural orthostatic tachycardia syndrome. The girls with orthostatic intolerance and CRPS commonly experienced transient violent tremors and persistent asthenia. Electron-microscopic examinations of the intradermal nerves showed an abnormal pathology in the unmyelinated fibers in two of the three girls examined. CONCLUSION: The symptoms observed in this study can be explained by abnormal peripheral sympathetic responses. The most common previous diagnosis in the studied girls was psychosomatic disease. The social problems of the study participants remained unresolved in that the severely disabled girls stopped going to school.
Asunto(s)
Enfermedades del Sistema Nervioso Autónomo/inducido químicamente , Inmunización/efectos adversos , Infecciones por Papillomavirus/prevención & control , Vacunas contra Papillomavirus/efectos adversos , Sistema Nervioso Simpático/fisiopatología , Adolescente , Enfermedades del Sistema Nervioso Autónomo/diagnóstico , Enfermedades del Sistema Nervioso Autónomo/epidemiología , Biopsia , Niño , Femenino , Estudios de Seguimiento , Humanos , Hipotensión Ortostática/inducido químicamente , Hipotensión Ortostática/diagnóstico , Hipotensión Ortostática/epidemiología , Incidencia , Japón/epidemiología , Pronóstico , Estudios Retrospectivos , Sistema Nervioso Simpático/efectos de los fármacosRESUMEN
Histological evaluation of a peripheral nerve is often the final diagnostic work-up for a neuropathy of unknown origin, and a distal sensory nerve is usually biopsied. Here, we report the case of a female patient with painful unilateral neuropathy in the upper arm. According to the histological evaluation of the pronator teres motor branch, vasculitis seemed to be the most probable cause of the condition, and steroid therapy improved the patients' symptoms. A biopsy of the motor branch of the pronator teres muscle nerve may be considered a valuable diagnostic option in selected cases with neuropathy affecting the upper limb, when performed in cooperation with neurologists and orthopedic surgeons.
RESUMEN
We report a patient with rheumatoid arthritis (RA) who developed leukoencephalopathy while being treated with low-dose methotrexate (MTX). She suddenly developed loss of recent memory and left homonymous hemianopsia ascribable to the bilateral but right-predominant occipitotemporal lesions. Intravenous administration of dexamethasone and cessation of MTX quickly relieved her clinical symptoms. Low-dose MTX-induced leukoencephalopathy is a rare complication in RA, but is important with regard to the possibility of serious neurological sequellae.
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Antirreumáticos/efectos adversos , Artritis Reumatoide/tratamiento farmacológico , Leucoencefalopatías/inducido químicamente , Metotrexato/efectos adversos , Anciano , Antirreumáticos/administración & dosificación , Dexametasona/uso terapéutico , Femenino , Humanos , Leucoencefalopatías/diagnóstico , Leucoencefalopatías/tratamiento farmacológico , Imagen por Resonancia Magnética , Metotrexato/administración & dosificaciónRESUMEN
Calciphylaxis is a vascular calcification-cutaneous necrosis syndrome, usually seen in patients with end-stage renal disease and secondary hyperparathyroidism. We report a 57-year-old polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome patient complicated with extensive skin ulcers due to calciphylaxis. He first noted a painful cutaneous ulcer on his left thigh, and then skin lesions rapidly worsened, resulting in multiple intractable ulcers with gangrene on his legs and trunk in a few months. Serum vascular endothelial growth factor (VEGF) was markedly elevated. Biopsy samples from his skin ulcers showed the deposition of calcium in the medial layer of cutaneous vessels, this finding being compatible with calciphylaxis. This is the second reported case with POEMS syndrome complicated with calciphylaxis. Both patients had no evidence of renal failure, hyperparathyroidism, or clotting disorders. The pathogenic link between POEMS syndrome and calciphylaxis is still unclear, but VEGF is known to regulate vascular calcification, in cooperation with bone morphogenetic proteins. Further, corticosteroid and several proinflammatory cytokines activate nuclear factor-κB pathway, known as the final common pathway leading to vascular calcification. Taken together, we consider that POEMS syndrome can be an independent risk condition for calciphylaxis.