ABSTRACT
Multiple sclerosis (MS) is an immune-mediated chronic central nervous system (CNS) disease affecting more than 400 000 people in the United States. Myelin-reactive CD4 T cells play critical roles in the formation of acute inflammatory lesions and disease progression in MS and experimental autoimmune encephalomyelitis (EAE), a well-defined mouse model for MS. Current MS therapies are only partially effective, making it necessary to develop more effective therapies that specifically target pathogenic myelin-specific CD4 T cells for MS treatment. While suppressing T-bet, the key transcription factor in T helper type 1 (Th1) cells, has been demonstrated to be beneficial in prevention and treatment of EAE, the therapeutic potential of retinoic acid-related orphan receptor gamma t (ROR)γt, the key transcription factor for Th17 cells, has not been well-characterized. In this study, we characterized the correlation between RORγt expression and other factors affecting T cell encephalitogenicity and evaluated the therapeutic potential of targeting RORγt by siRNA inhibition of RORγt. Our data showed that RORγt expression correlates with interleukin (IL)-17 production, but not with the encephalitogenicity of myelin-specific CD4 T cells. IL-23, a cytokine that enhances encephalitogenicity, does not enhance RORγt expression significantly. Additionally, granulocyte-macrophage colony-stimulating factor (GM-CSF) levels, which correlate with the encephalitogenicity of different myelin-specific CD4 T cell populations, do not correlate with RORγt. More importantly, inhibiting RORγt expression in myelin-specific CD4 T cells with an siRNA does not reduce disease severity significantly in adoptively transferred EAE. Thus, RORγt is unlikely to be a more effective therapeutic target for ameliorating pathogenicity of encephalitogenic CD4 T cells.
Subject(s)
Central Nervous System/immunology , Central Nervous System/metabolism , Nuclear Receptor Subfamily 1, Group F, Member 3/genetics , Animals , CD4-Positive T-Lymphocytes/drug effects , CD4-Positive T-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/metabolism , Disease Models, Animal , Encephalomyelitis, Autoimmune, Experimental/genetics , Encephalomyelitis, Autoimmune, Experimental/immunology , Gene Expression , Gene Expression Regulation , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Interleukin-23/metabolism , Interleukin-23/pharmacology , Mice , Mice, Knockout , Multiple Sclerosis/genetics , Multiple Sclerosis/immunology , Myelin Sheath/immunology , Nuclear Receptor Subfamily 1, Group F, Member 3/metabolism , RNA InterferenceABSTRACT
Apoptosis induced by tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL/APO-2L) has been shown to exert important functions during various immunological processes. The involvement of the death adaptor proteins FADD/MORT1, TRADD, and RIP and the apoptosis-initiating caspases-8 and -10 in death signaling by the two death-inducing TRAIL receptors 1 and 2 (TRAIL-R1 and TRAIL-R2) are controversial. Analysis of the native TRAIL death-inducing signaling complex (DISC) revealed ligand-dependent recruitment of FADD/MORT1 and caspase-8. Differential precipitation of ligand-stimulated TRAIL receptors demonstrated that FADD/MORT1 and caspase-8 were recruited to TRAIL-R1 and TRAIL-R2 independently of each other. FADD/MORT1- and caspase-8-deficient Jurkat cells expressing only TRAIL-R2 were resistant to TRAIL-induced apoptosis. Thus, FADD/MORT1 and caspase-8 are essential for apoptosis induction via TRAIL-R2.
Subject(s)
Adaptor Proteins, Signal Transducing , Apoptosis/immunology , Carrier Proteins/physiology , Caspases/physiology , Receptors, Tumor Necrosis Factor/physiology , fas Receptor/physiology , B-Lymphocytes/cytology , B-Lymphocytes/enzymology , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Carrier Proteins/metabolism , Caspase 8 , Caspase 9 , Caspases/metabolism , Cell Line , Fas-Associated Death Domain Protein , Humans , Jurkat Cells , Receptors, TNF-Related Apoptosis-Inducing Ligand , Receptors, Tumor Necrosis Factor/biosynthesis , Signal Transduction/immunology , T-Lymphocytes/cytology , T-Lymphocytes/enzymology , T-Lymphocytes/immunology , T-Lymphocytes/metabolism , Tumor Cells, CulturedABSTRACT
The seeds of Guaraná are rich in xanthines and are used for the preparation of guaraná powder which is very commonly given to horses as a 'tonic' in Brazil. In this paper, the xanthine content of guaraná powder was determined, in addition to its clearance time in horses. Thin-layer chromatography was used as a screening procedure and high-performance liquid chromatography was performed to quantify the drugs in both the powder and urine samples. The guaraná powder was found to contain 2.16, 1.10 and 36.78 mg g-1 of theobromine (TB), theophylline (TP) and caffeine (CF), respectively, and in urine it was possible to detect TB and TP up to 13 d and CF up to 9 d after the administration of guaraná powder.
Subject(s)
Caffeine/administration & dosage , Theobromine/administration & dosage , Theophylline/administration & dosage , Xanthines/urine , Administration, Oral , Animals , Caffeine/analysis , Caffeine/chemistry , Caffeine/metabolism , Chromatography, High Pressure Liquid/methods , Chromatography, Thin Layer/methods , Drug Combinations , Horses , Regression Analysis , Reproducibility of Results , Seeds , Theobromine/analysis , Theobromine/chemistry , Theobromine/metabolism , Theophylline/analysis , Theophylline/chemistry , Theophylline/metabolism , Xanthines/analysisABSTRACT
The troublesome symptoms of the seriously defective schizophrenics depend sometimes upon their unexpected sensitive experiences, which are not reported verbally and are guessed merely through the precise observations of the behaviour. By the treatment of the disturbances so found, the troublesome symptoms disappear. As an example of this "behaviour-analytical treatment" an old, defective schizophrenic with incontinence and negativism was introduced in this report, whose background suggested an intensive mysophobia, and was treated successfully. Some forsaken inmates of the asylums might be helped in this way.