ABSTRACT
We report a singular case of pigmented pagetoid Bowen's disease showing transitional features between extramammary Paget's disease and in situ squamous cell carcinoma.^ieng
Differentiation of pagetoid cutaneous neoplasms can be very challenging on hematoxylin and eosin-stained sections.
Subject(s)
Adenocarcinoma in Situ/diagnosis , Bowen's Disease/diagnosis , Skin Neoplasms/diagnosis , Adenocarcinoma in Situ/pathology , Aged , Bowen's Disease/pathology , Diagnosis, Differential , Female , Humans , Skin Neoplasms/pathologyABSTRACT
STUDY DESIGN: Cell transplantation strategies are gaining increasing interest for spinal cord injury (SCI) with the objective of promoting spinal cord repair. To avoid allogenic graft rejection, an adequate immune suppression is required, and one of the most potent and commonly used immunosuppressives is cyclosporin A (CsA). In SCI, permanent sensory motor loss is combined with modifications of drug absorption, distribution and elimination. OBJECTIVES: The objectives of this study were to thoroughly explore histological and functional outcomes of CsA treatment in a rat model of spinal cord compression. SETTING: Experiments were carried out at the Institute for Neurosciences of Montpellier (France), the Integrative Biology of Neurodegeneration Laboratory (Spain) and in the Novartis Institutes for BioMedical Research (Switzerland) for CsA blood concentration determination. METHODS: We first evaluated histological outcomes of CsA treatment on kidneys and spinal cord after SCI. We then investigated whether SCI modified CsA blood concentration. Finally, using behavioral analysis, we assessed the potential CsA impact on functional recovery. RESULTS: When spinal-cord-injured rats were treated with a CsA dose of 10 mg kg(-1) per day, we observed deleterious effects on kidneys, associated with modifications of CsA blood concentration. Adding an antibiotic treatment reduced kidney alteration without modifying CsA blood concentration. Finally, we showed that CsA treatment per se modified neither functional recovery nor lesion extension. CONCLUSION: This study pinpoints the absolute requirement of careful CsA monitoring in the clinical setting for patients with SCI to minimize potential unexpected effects and avoid therapeutic failure.
Subject(s)
Cell Transplantation/methods , Cyclosporine/toxicity , Graft Rejection/drug therapy , Kidney Diseases/physiopathology , Spinal Cord Injuries/physiopathology , Animals , Cell Transplantation/adverse effects , Cyclosporine/blood , Cyclosporine/therapeutic use , Disease Models, Animal , Drug Therapy, Combination/methods , Graft Rejection/physiopathology , Graft Rejection/prevention & control , Immunosuppressive Agents/blood , Immunosuppressive Agents/therapeutic use , Immunosuppressive Agents/toxicity , Kidney Diseases/chemically induced , Male , Rats , Rats, Wistar , Spinal Cord Injuries/surgery , Treatment OutcomeABSTRACT
Astrocytes play an active role in the brain and spinal cord. For example, they have a function in formation and maintenance of the blood-brain barrier, ion homeostasis, neurotransmitter transport, production of extracellular matrix, and neuromodulation. Moreover, they play a role in preserving or even restoring the structural and physiological integrity after tissue injury. Currently, the function of astrocytes was studied with regard to the controversially discussed aspects of permissivity on the one-hand-side and inhibition of the other side exerted by reactive astrocytes for axonal regrowth in the adult CNS. Accordingly, knock-out mice deficient in vimentin (VIM) and/or glial fibrillary acidic protein (GFAP), the two major IF-proteins of astrocytes, were investigated. In addition, in vitro studies were carried out, on whether the absence of one or both proteins (VIM, GFAP) influences axonal regeneration. In experimental animals, a hemisection of the spinal cord was performed utilizing the above mentioned double-mutant mice. The knock-out mice were generated by gene targeting. Double-mutants were obtained by crossing single null mice. The in vitro results indicate that both VIM and GFAP were absent in astrocytic cultures obtained from double-mutant mice. On the other side, the proteins were detected in more than 85%, of cultured cells from wild types. Co-culture of mutant mice astrocytes with neurons revealed that the neuronal density was different from that obtained in culture with wild type astrocytes. On the other side, there was a marked increase in neuronal density in co-cultures utilizing both GFAP knock-out- or double-mutant mice astrocytes again as compared to co-cultures with wild type astrocytes. Moreover, the neurite length of neurons was significantly increased in experiments with neurons growing on astrocytes from GFAP-knock-out or double-mutant mice. The in vivo experiments demonstrate an increase of nestin (NES) immunoreactivity at three days in the sectioned side of the spinal cord, in the perikaryon and astroglial processes. In double-mutant mice only a slight increase in NES-immunoreactivity was found in the lesion side, albeit confined to the perikaryon of astrocytes. Below the lesion, serotonin immunostaining was dramatically reduced three days after the insult in both sides, particularly in the lesion side. The decrease was more pronounced in double-mutant than in wild type mice. On the other side, double-mutant mice had a much higher density of serotonergic fibers in the ventral horn in the lesioned side. In conclusion, the findings demonstrate that in the absence of important astrocytic proteins as VIM and GFAP, the astroglial response to injury is significantly modified underlying reduced scar formation. Attenuation of scar formation may enhance axonal sprouting of serotonergic axons below the lesion, which specifically reinnervate motoneuron pools.
Subject(s)
Astrocytes/pathology , Axons/pathology , Glial Fibrillary Acidic Protein/deficiency , Nerve Regeneration , Neurons/pathology , Spinal Cord Injuries/pathology , Spinal Cord Injuries/physiopathology , Vimentin/deficiency , Animals , Cells, Cultured , Central Nervous System/pathology , Central Nervous System/physiopathology , Coculture Techniques/methods , Glial Fibrillary Acidic Protein/genetics , Mice , Mice, Knockout , Mice, Transgenic , Neuroglia/pathology , Vimentin/geneticsABSTRACT
A great deal of interest has attracted the attention of researchers on the potential use of (neural) stem cells in cell replacement or restorative therapies for heretofore incurable CNS pathologies such as brain stroke, spinal cord injury, Parkinson's disease or multiple sclerosis. This short perspective illustrates our view of neural stem cell research with a focus on the stem cell concept, on the in situ identity of neural stem cells and on selected aspects of embryonic and adult neurogenesis. A brief survey of current stem cell-based experimental literature tries to provide a realistic picture of how far we have gone in the quest to establish a restorative neurology.
Subject(s)
Neurology , Neurons/cytology , Stem Cells/cytology , Animals , Cell Differentiation , Humans , Multiple Sclerosis/therapy , Parkinson Disease/therapy , Research , Spinal Cord Injuries/therapy , Stroke/therapyABSTRACT
The lack of axonal regeneration in the injured adult mammalian spinal cord leads to permanent functional disabilities. The inability of neurons to regenerate their axon is appreciably due to an inhospitable environment made of an astrocytic scar. We generated mice knock-out for glial fibrillary acidic protein and vimentin, the major proteins of the astrocyte cytoskeleton, which are upregulated in reactive astrocytes. These animals, after a hemisection of the spinal cord, presented reduced astroglial reactivity associated with increased plastic sprouting of supraspinal axons, including the reconstruction of circuits leading to functional restoration. Therefore, improved anatomical and functional recovery in the absence of both proteins highlights the pivotal role of reactive astrocytes in axonal regenerative failure in adult CNS and could lead to new therapies of spinal cord lesions.
Subject(s)
Glial Fibrillary Acidic Protein/deficiency , Spinal Cord Injuries/physiopathology , Vimentin/deficiency , Animals , Astrocytes/physiology , Axons/physiology , Female , Glial Fibrillary Acidic Protein/genetics , Glial Fibrillary Acidic Protein/physiology , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mice, Knockout , Motor Activity , Nerve Regeneration , Neuronal Plasticity , Spinal Cord Injuries/pathology , Spinal Cord Injuries/therapy , Vimentin/genetics , Vimentin/physiologyABSTRACT
One kidney was taken from each of 100 pigs at slaughter; 50 had gross lesions of multifocal interstitial nephritis and 50 had no gross lesions. Forty-nine of the affected kidneys had lesions that were characterised by the presence of either a few randomly distributed or numerous widely disseminated pale foci, 1 to 3 mm in diameter, on the cortical surface (white-dotted kidneys). Microscopically, these focal inflammatory lesions often had a distinct lymphofollicular pattern (follicular nephritis). Lesions of chronic vasculitis were observed in 21 of the affected kidneys. Histologically, the control kidneys had only small and sparse inflammatory foci. Standard bacterial cultures of kidneys of both groups were not significant, and cultures for the isolation of leptospires were all negative. Virological examination of the kidney homogenates by PCR did not reveal any porcine reproductive and respiratory syndrome virus and only a few cases were positive for the porcine circovirus type 1. However, porcine parvovirus (PPV) and porcine circovirus type 2 (PCV-2) were detected in many kidneys of both groups but in a significantly higher proportion of the kidneys with interstitial nephritis. There was a significant association between the lesions and the presence of PPV and PCV-2 with odds ratios of 7.5 (P<0.0001) and 3.4 (P=0.0074), respectively, and the odds ratio increased to 22.7 (P<0.0001) when both viruses were identified in the same kidney. However, a subsample of kidneys taken from both groups were negative by immunohistochemistry for the presence of PPV and PCV-2 antigens.
Subject(s)
Nephritis, Interstitial/veterinary , Swine Diseases/pathology , Animals , Nephritis, Interstitial/microbiology , Nephritis, Interstitial/pathology , Polymerase Chain Reaction , Swine , Swine Diseases/microbiologyABSTRACT
Treatment of neurodegenerative diseases by classical pharmacotherapy is restricted by blood-brain barrier which prevents access to the brain of potentially therapeutic molecules. Recent progress in the knowledge of pathophysiological molecular processes, and in the development of molecular biotechnology have opened the way to new therapeutic interventions for these disorders. This chapter reviews the most recent gene therapy strategies using experimental models for neurodegenerative diseases.
Subject(s)
Genetic Therapy/methods , Neurodegenerative Diseases/therapy , Alzheimer Disease/therapy , Amyotrophic Lateral Sclerosis/therapy , Animals , Deafness/therapy , Genetic Vectors , Humans , Huntington Disease/therapy , Neurons , Parkinson Disease/therapy , Parkinsonian Disorders/therapy , Promoter Regions, Genetic , Retinitis Pigmentosa/therapyABSTRACT
Intermediate filaments (IFs) are a major component of the cytoskeleton in astrocytes. Their role is far from being completely understood. Immature astrocytes play a major role in neuronal migration and neuritogenesis, and their IFs are mainly composed of vimentin. In mature differentiated astrocytes, vimentin is replaced by the IF protein glial fibrillary acidic protein (GFAP). In response to injury of the CNS in the adult, astrocytes become reactive, upregulate the expression of GFAP, and reexpress vimentin. These modifications contribute to the formation of a glial scar that is obstructive to axonal regeneration. Nevertheless, astrocytes in vitro are considered to be the ideal substratum for the growth of embryonic CNS axons. In the present study, we have examined the potential role of these two major IF proteins in both neuronal survival and neurite growth. For this purpose, we cocultured wild-type neurons on astrocytes from three types of knock-out (KO) mice for GFAP or/and vimentin in a neuron-astrocyte coculture model. We show that the double KO astrocytes present many features of immaturity and greatly improve survival and neurite growth of cocultured neurons by increasing cell-cell contact and secreting diffusible factors. Moreover, our data suggest that the absence of vimentin is not a key element in the permissivity of the mutant astrocytes. Finally, we show that only the absence of GFAP is associated with an increased expression of some extracellular matrix and adhesion molecules. To conclude, our results suggest that GFAP expression is able to modulate key biochemical properties of astrocytes that are implicated in their permissivity.
Subject(s)
Glial Fibrillary Acidic Protein/biosynthesis , Nerve Tissue Proteins , Neural Cell Adhesion Molecules/biosynthesis , Neurites/metabolism , Neurons/metabolism , Vimentin/biosynthesis , Animals , Astrocytes/metabolism , Astrocytes/ultrastructure , Cadherins/metabolism , Cell Adhesion/physiology , Cell Count , Cell Survival/genetics , Cells, Cultured , Coculture Techniques , Extracellular Matrix/metabolism , Fibronectins/metabolism , Gene Targeting , Glial Fibrillary Acidic Protein/deficiency , Glial Fibrillary Acidic Protein/genetics , Intermediate Filament Proteins/biosynthesis , Intermediate Filaments/metabolism , Intermediate Filaments/ultrastructure , Mice , Mice, Inbred Strains , Mice, Knockout , Models, Biological , Nestin , Neural Cell Adhesion Molecules/metabolism , Neurites/ultrastructure , Neurons/cytology , Vimentin/deficiency , Vimentin/geneticsABSTRACT
During the development of the CNS, astrocytes play a key role as a substrate for neuronal migration and axonal growth. These neuron-astrocyte interactions could be regulated, in part, by the astrocytic cytoskeleton. Nestin, vimentin, and glial fibrillary acidic protein (GFAP) are the three identified proteins constitutive of intermediate filaments present in astrocytes. In the present study, we used mice deficient in GFAP to define the influence of the major protein of the astrocytic cytoskeleton on neuron survival and axonal growth in a model of neuron-astrocyte coculture. We observed that GFAP null astrocytes are a better substrate for neuronal survival and neurite outgrowth than wild-type astrocytes. This may be correlated with the relatively late occurrence of GFAP expression in astrocyte maturation when the early steps of neurogenesis are completed.
Subject(s)
Astrocytes/metabolism , Cell Survival/physiology , Central Nervous System/embryology , Glial Fibrillary Acidic Protein/deficiency , Nerve Tissue Proteins , Neurites/metabolism , Neurons/metabolism , Animals , Astrocytes/ultrastructure , Cells, Cultured , Central Nervous System/metabolism , Central Nervous System/ultrastructure , Cytoskeleton/metabolism , Cytoskeleton/ultrastructure , Fetus , Glial Fibrillary Acidic Protein/genetics , Growth Cones/metabolism , Growth Cones/ultrastructure , Intermediate Filament Proteins/metabolism , Mice , Mice, Knockout , Nestin , Neurites/ultrastructure , Neurons/ultrastructure , Vimentin/metabolismSubject(s)
Acute Kidney Injury/veterinary , Dog Diseases/microbiology , Leptospira/pathogenicity , Leptospirosis/veterinary , Acute Kidney Injury/etiology , Acute Kidney Injury/microbiology , Animals , Cross Reactions , Dog Diseases/immunology , Dogs , Leptospira/immunology , Leptospirosis/diagnosis , Leptospirosis/immunology , Sensitivity and Specificity , Serologic Tests/veterinary , Specimen HandlingABSTRACT
Traumatic lesions of the spinal cord yield a loss of supraspinal control of voluntary locomotor activity, although the spinal cord contains the necessary circuitry to generate the basic locomotor pattern. In spinal rats, this network, known as central pattern generator (CPG), was shown to be sensitive to serotonergic pharmacological stimulation. In previous works we have shown that embryonic raphe cells transplanted into the sublesional cord of adult rats can reinnervate specific targets, restore the lesion-induced increase in receptor densities of neurotransmitters, promote hindlimb weight support, and trigger a locomotor activity on a treadmill without any other pharmacological treatment or training. With the aim of discriminating whether the action of serotonin on CPG is associated to a specific level of the cord, we have transplanted embryonic raphe cells at two different levels of the sublesional cord (T9 and T11) and then performed analysis of the kinematic and EMG activity synchronously recorded during locomotion. Locomotor performances were correlated to the reinnervated level of the cord and compared to that of intact and transected nontransplanted animals. The movements expressed by T11 transplanted animals correspond to a well defined locomotor pattern comparable to that of the intact animals. On the contrary, T9 transplanted animals developed limited and disorganized movements as those of nontransplanted animals. The correlation of the locomotor performances with the level of reinnervation of the spinal cord suggests that serotonergic reinnervation of the L1-L2 level constitutes a key element in the genesis of this locomotor rhythmic activity. This is the first in vivo demonstration that transplanted embryonic raphe cells reinnervating a specific level of the cord activate a locomotor behavior.
Subject(s)
Brain Tissue Transplantation/physiology , Fetal Tissue Transplantation/physiology , Locomotion/physiology , Motor Activity/physiology , Raphe Nuclei/cytology , Spinal Cord Injuries/physiopathology , Spinal Cord Injuries/surgery , Spinal Cord/physiopathology , Animals , Biomechanical Phenomena , Electromyography , Female , Lumbar Vertebrae , Muscle, Skeletal/innervation , Rats , Rats, Sprague-Dawley , Spinal Cord/physiologyABSTRACT
In the cerebellum of adult mammals, glial fibrillary acidic protein (GFAP) and vimentin (VIM) are coexpressed in Golgi epithelial cells (GEC), also known as Bergmann glia. In this study we used three transgenic knockout mice (GFAP, VIM and double GFAP and VIM) to analyze the involvement of these proteins in the building of glial filaments and in neuron-glia interactions. The cerebella of VIM, GFAP, and GFAP/VIM mutant mice were processed by the rapid Golgi method and also for electron microscopy. In VIM mutant mice, Bergmann fibers are hypertrophic with thickened appendages. In the electron microscope they appear as large glial profiles devoid of glial filaments, with embedded dendritic thorns and parallel fiber boutons. In addition, signs of degeneration are observed in Purkinje cells. In GFAP mutant mice, GEC exhibit fine, delicate processes, as those seen in wild-type animals, however, a large accumulation of lamellae and granular appendages was observed along their surfaces, which came into contact with each other. The electron microscope exhibited fine and scarce astroglial profiles containing some glial filaments, a stunted glia limitans, and the presence of large extracellular spaces. In double mutant mice, the two phenotypes are expressed but appear attenuated, with a total absence of glial filaments and the general appearance of immaturity for GEC. In conclusion, it appears that the absence of each of the proteins yields a specific phenotype and that the defects are not necessarily additive.
Subject(s)
Cerebellar Cortex/pathology , Glial Fibrillary Acidic Protein/deficiency , Vimentin/deficiency , Animals , Glial Fibrillary Acidic Protein/genetics , Mice , Mice, Knockout/anatomy & histology , Mice, Knockout/genetics , Microscopy, Electron , Vimentin/geneticsABSTRACT
Serology plays an important role in the diagnosis of leptospirosis. Few laboratories have the resources, expertise, or facilities to perform the microscopic agglutination test (MAT). Thus, there is a need for a rapid and simple serological test that could be used in any diagnostic laboratory. In this study, a genus-specific, heat-stable antigenic preparation from Leptospira interrogans serovar pomona was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of leptospiral antibodies in dog sera. This antigenic preparation reacted with rabbit antisera against L. interrogans serovars bratislava, autumnalis, icterohaemorrhagiae and pomona and with rabbit antiserum against L. kirschneri serovar grippotyphosa. The ELISA showed a relative specificity of 95.6% with 158 dog sera which were negative at a dilution of 1:100 in the MAT for serovars pomona, bratislava, icterohaemorrhagiae, autumnalis, hardjo, and grippotyphosa. The relative sensitivity of this assay with 21 dog sera that revealed serovars MAT titres of > or =100 to different serovars was 100%. This assay is easily standardized, technically more advantageous than MAT, and uses an antigenic preparation that can be routinely prepared in large amounts. It was concluded that this ELISA is sufficiently sensitive test to be used as an initial screening test for the detection of leptospiral antibodies in canine sera, with subsequent confirmation of positive test results with the MAT.
Subject(s)
Antibodies, Bacterial/analysis , Dog Diseases/microbiology , Enzyme-Linked Immunosorbent Assay/veterinary , Leptospira/immunology , Leptospirosis/veterinary , Animals , Dog Diseases/diagnosis , Dogs , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/standards , Leptospirosis/diagnosis , Rabbits , Sensitivity and SpecificityABSTRACT
Transection of the spinal cord yields a permanent deficit due to the interruption of descending and ascending tracts which subserve the supraspinal control of spinal cord functions. We have shown previously that transplantation below the level of the section of embryonic monoaminergic neurons can promote the recovery of some segmental functions via a local serotonergic and noradrenergic reinnervation. Moreover, the up-regulation of the corresponding receptors resulting from the section was corrected by the transplants. The aim of the present work was to determine whether such a graft could also influence non-monoaminergic local neurons, the GABAergic interneurons of the spinal cord. Following spinal cord transection, the number of cells which express glutamate decarboxylase (mol. wt 67,000) messenger RNA--a marker of GABA synthesis--increased significantly below the lesion compared with the intact animal. In contrast, in lesioned animals which had been grafted one week later with raphe neuroblasts, this number was close to control level. These post-grafting modifications were further associated with increased GABA immunoreactivity in the host tissue. These data suggest that the graft of embryonic raphe cells which compensates the deficit of serotonin in the distal segment also regulates the expression of the GABAergic phenotype in the host spinal cord. This regulation could be mediated by the re-establishment of a local functional innervation by both serotonin and GABAergic transplanted neurons and/or by trophic factors released from the embryonic cells. It appears then that grafted cells influence the host tissue in a complex manner, through the release and/or regulation of several neurotransmitter systems.
Subject(s)
Cell Transplantation , Fetal Tissue Transplantation , Raphe Nuclei/embryology , Spinal Cord Injuries/physiopathology , Spinal Cord Injuries/surgery , gamma-Aminobutyric Acid/metabolism , Animals , Embryo, Mammalian/cytology , Female , Immunohistochemistry , In Situ Hybridization , Interneurons/physiology , Phenotype , Raphe Nuclei/cytology , Rats , Rats, Sprague-Dawley , Spinal Cord/metabolism , Spinal Cord Injuries/genetics , Spinal Cord Injuries/pathologyABSTRACT
To investigate the characteristics of neurotropic signaling involved in specific target recognition by grafted embryonic serotonergic cells, we have developed an in vitro grafting model. Specific raphe nuclei (B1/B2 and B3) were respectively dissected from 14-day-old rat embryos, and partially dissociated cells were cocultured on spinal cord slices from 20-day-old fetuses. After serotonin immunodetection, neurite growth patterns were analyzed by standard photonic or confocal scanning microscopy. Computer reconstruction (maximal signal projection) was used to track individual neurites in spite of their changing depth levels. Whereas the direction and branching of the initial neurite segments did not seem to be significantly influenced by any specific environment, specific growth patterns were developed at some distance from the cell bodies, indicating that neurites are able to recognize their specific targets.
Subject(s)
Neurons/physiology , Raphe Nuclei/physiology , Spinal Cord/physiology , Animals , Coculture Techniques , Immunohistochemistry , Microscopy, Confocal , Neurites/metabolism , Neurites/physiology , Neurons/transplantation , Raphe Nuclei/cytology , Raphe Nuclei/embryology , Rats , Rats, Sprague-Dawley , Serotonin/metabolism , Spinal Cord/embryologyABSTRACT
The aim of this study was to assess the biological characteristics of four new malignant mesothelioma (MM) cell lines. Since simian virus (SV)40 sequences have been recently detected in MM, SV40 large T antigen (Tag) expression was also analysed. MM cell lines were characterized by morphological, ultrastructural and cytogenetic analysis. Expression of Tag and of relevant MM markers was studied by immunocytochemistry, surface antigens by indirect immunofluorescence and immunomodulating cytokines by enzyme-linked immunosorbent assay (ELISA). The four MM cell lines, established from pleural effusions, showed a slow proliferation rate and pleomorphic changes during culture. Cell lines expressed vimentin, cytokeratins 8 and 18, and the mesothelial antigen recognized by HBME-1 monoclonal antibody, but not carcinoembryonic antigen. Surface human leukocyte antigen (HLA)-class I and intercellular adhesion molecule (ICAM)-1 molecules were present on all the cell lines. While HLA class II and CD86 were constitutively undetectable, HLA-class II was present after interferon (IFN)-gamma stimulation. All cell lines displayed abnormal karyotypes with chromosome 6 abnormalities. Transforming growth factor (TGF)-beta2 and interleukin (IL)-6 were constitutively secreted, while tumour necrosis factor (TNF)-alpha was secreted only in response to lipopolysaccharide. Intranuclear Tag was expressed in two cell lines. The persistence of large T antigen with human leukocyte antigen class I and intercellular adhesion molecule-1 positivity may point to large T antigen as a target for cytotoxic T-lymphocyte-based immunotherapy in some malignant mesothelioma patients.