ABSTRACT
According to one hypothesis, Parkinson's disease pathogenesis is largely caused by dopamine catabolism that is catalyzed on mitochondrial membranes by monoamine oxidase. Reactive oxygen species are formed as a byproduct of these reactions, which can lead to mitochondrial damage followed by cell degeneration and death. In this study, we investigated the effects of administration of the mitochondrial antioxidant SkQ1 on biochemical, immunohistochemical, and behavioral parameters in a Parkinson-like condition caused by protoxin MPTP injections in C57BL/6 mice. SkQ1 administration increased dopamine quantity and decreased signs of sensory-motor deficiency as well as destruction of dopaminergic neurons in the substantia nigra and ventral tegmental area in mice with the Parkinson-like condition.
Subject(s)
Antioxidants/therapeutic use , MPTP Poisoning/drug therapy , Mitochondria/metabolism , Plastoquinone/analogs & derivatives , 3,4-Dihydroxyphenylacetic Acid/analysis , Animals , Antioxidants/pharmacology , Brain/metabolism , Brain/pathology , Dihydroxyphenylalanine/analysis , Dopamine/analysis , Immunohistochemistry , MPTP Poisoning/pathology , Maze Learning/drug effects , Mice , Mice, Inbred C57BL , Mitochondria/drug effects , Motor Activity/drug effects , Plastoquinone/pharmacology , Plastoquinone/therapeutic use , Tyrosine 3-Monooxygenase/metabolismABSTRACT
An effective system for expression of human granulocyte and granulocyte macrophage colony-stimulating factors (hG-CSF and hGM-CSF) in Nicotiana benthamiana plants was developed using viral vector based on tobacco mosaic virus infecting cruciferous plants. The genes of target proteins were cloned into the viral vector driven by actin promoter of Arabidopsis thaliana. The expression vectors were delivered into plant cells by agroinjection. Maximal synthesis rate was detected 5 days after injection and was up to 500 and 300 mg per kg of fresh leaves for hG-CSF and hGM-CSF, respectively. The yield of purified hG-CSF and hGM-CSF was 100 and 50 mg/kg of fresh leaves, respectively. Recombinant plant-made hG-CSF and hGM-CSF stimulated proliferation of murine bone marrow and human erythroleucosis TF-1 cells, respectively, at the same rate as the commercial drugs.
Subject(s)
Genetic Engineering/methods , Granulocyte Colony-Stimulating Factor/metabolism , Granulocyte Colony-Stimulating Factor/pharmacology , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Nicotiana/genetics , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cells, Cultured , Gene Expression , Granulocyte Colony-Stimulating Factor/genetics , Granulocyte Colony-Stimulating Factor/isolation & purification , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Granulocyte-Macrophage Colony-Stimulating Factor/isolation & purification , Humans , Mice , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Recombinant Proteins/pharmacology , Nicotiana/metabolismABSTRACT
Escherichia coli bacterial cells of two strains JM109 and K12 J62 were imaged with atomic force microscopy (AFM) in different environmental conditions. The AFM results show that the two strains have considerable difference in the surface morphology. At the same time after rehydration both strains show the loss of the topographic features and increase in lateral and vertical dimensions. Results obtained in different AFM modes (contact, tapping, MAC) were compared. Imaging in culture medium was applied for direct observation of the surface degradation effect of lysozyme. The treatment of the cells with the enzyme in the culture medium lead to the loss of surface rigidity and eventually to dramatic changes of the bacteria shape.
Subject(s)
Escherichia coli/physiology , Escherichia coli/ultrastructure , Microscopy, Atomic Force/methods , Air Microbiology , Culture Media , Muramidase/metabolism , Surface Properties , Water , Water MicrobiologyABSTRACT
Plant virus-encoded movement proteins promote viral spread between plant cells via plasmodesmata. The movement is assumed to require a plasmodesmata targeting signal to interact with still unidentified host factors presumably located on plasmodesmata and cell walls. The present work indicates that a ubiquitous cell wall-associated plant enzyme pectin methylesterase of Nicotiana tabacum L. specifically binds to the movement protein encoded by tobacco mosaic virus. We also show that pectin methylesterase is an RNA binding protein. These data suggest that pectin methylesterase is a host cell receptor involved in cell-to-cell movement of tobacco mosaic virus.
