RESUMEN
AIMS: Our main objective was to determine the neuropathological correlates of dementia in patients with Lewy body disease (LBD). Furthermore, we used data derived from clinical, neuropathological and genetic studies to investigate boundary issues between Dementia with Lewy bodies (DLB) and Parkinson's disease with (PDD) and without (PDND) dementia. METHODS: One hundred and twenty-one cases with a neuropathological diagnosis of LBD and clinical information on dementia status were included in the analysis (55 PDD, 17 DLB and 49 PDND). We carried out topographical and semi-quantitative assessment of Lewy bodies (LB), Aß plaques and tau-positive neuropil threads (NT). The APOE genotype and MAPT haplotype status were also determined. RESULTS: The cortical LB (CLB) burden was the only independent predictor of dementia (OR: 4.12, P < 0.001). The total cortical Aß plaque burden was an independent predictor of a shorter latency to dementia from onset of motor signs (P = 0.001). DLB cases had a higher LB burden in the parietal and temporal cortex, compared to PDD. Carrying at least one APOE ϵ4 allele was associated with a higher cortical LB burden (P = 0.02), particularly in the neocortical frontal, parietal and temporal regions. CONCLUSIONS: High CLB burden is a key neuropathological substrate of dementia in LBD. Elevated cortical LB pathology and Aß plaque deposition are both correlated with a faster progression to dementia. The higher CLB load in the temporal and parietal regions, which seems to be a distinguishing feature of DLB, may account for the shorter latency to dementia and could be mediated by the APOE ϵ4 allele.
Asunto(s)
Corteza Cerebral/patología , Demencia/epidemiología , Cuerpos de Lewy/patología , Enfermedad por Cuerpos de Lewy/patología , Enfermedad de Parkinson/patología , Adulto , Anciano , Anciano de 80 o más Años , Péptidos beta-Amiloides/metabolismo , Demencia/etiología , Demencia/patología , Femenino , Humanos , Enfermedad por Cuerpos de Lewy/complicaciones , Masculino , Persona de Mediana Edad , Enfermedad de Parkinson/complicaciones , PrevalenciaRESUMEN
UNLABELLED: During neuroinflammation in multiple sclerosis (MS) fibrinogen, not normally present in the brain or spinal cord, enters the central nervous system through a compromised blood-brain barrier. Fibrin deposited on axons is ineffectively removed by tissue plasminogen activator (tPA), a key contributory factor being the upregulation of plasminogen activator inhibitor-1 (PAI-1). AIMS: This study investigated the role of PAI-1 during experimental neuroinflammatory disease. METHODS: Chronic relapsing experimental allergic encephalomyelitis (CREAE), a model of MS, was induced with spinal cord homogenate in PAI-1 knockout (PAI-1(-/-)) and wild type (WT) mice, backcrossed onto the Biozzi background. RESULTS: Disease incidence and clinical severity were reduced in PAI-1(-/-) mice, with animals developing clinical signs significantly later than WTs. Clinical relapses were absent in PAI-1(-/-) mice and the subsequent reduction in neuroinflammation was coupled with a higher capacity for fibrinolysis in spinal cord samples from PAI-1(-/-) mice, in association with increased tPA activity. Axonal damage was less apparent in PAI-1(-/-) mice than in WTs, implicating fibrin in both inflammatory and degenerative events during CREAE. CONCLUSIONS: PAI-1 is a potential target for therapy in neuroinflammatory degenerative diseases, allowing effective fibrin removal and potentially reducing relapse rate and axonal damage.
Asunto(s)
Encefalomielitis Autoinmune Experimental/metabolismo , Encefalomielitis Autoinmune Experimental/patología , Fibrinólisis/fisiología , Inhibidor 1 de Activador Plasminogénico/metabolismo , Animales , Axones/metabolismo , Axones/patología , Western Blotting , Enfermedad Crónica , Encefalomielitis Autoinmune Experimental/inmunología , Ensayo de Inmunoadsorción Enzimática , Inmunohistoquímica , Inflamación/patología , Ratones , Ratones Noqueados , Médula Espinal/inmunología , Médula Espinal/metabolismo , Médula Espinal/patología , Activador de Tejido Plasminógeno/metabolismoRESUMEN
In multiple sclerosis (MS), the matrix metalloprotease (MMP) gelatinase B/MMP-9 and platelet endothelial cell adhesion molecule (PECAM)-1 have both been implicated in trans-endothelial infiltration of leucocytes into the brain, but their functional connection has not yet been investigated. We investigated the expression of gelatinase B and PECAM-1 in post mortem brains of MS patients by immunohistochemistry. Because increased soluble PECAM-1 serum levels have been observed in MS patients, we also tested in vitro whether this could be due to cleavage of PECAM-1 by gelatinase B or matrilysin-1/MMP-7. Constitutive expression of PECAM-1 was found on brain endothelial cells, whilst in active MS lesions cell-bound PECAM-1 was highly up-regulated on foamy macrophages in perivascular infiltrates and co-localized with gelatinase B. However, human THP-1 monocyte-bound or soluble recombinant PECAM-1 were both resistant to proteolytic cleavage by gelatinase B or matrilysin-1 in vitro, as demonstrated by Western blot analysis and flow cytometry. These results suggest that PECAM-1 and gelatinase B may complement each other during the transmigration of the blood-brain barrier by mononuclear cells.
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Encéfalo/irrigación sanguínea , Células Endoteliales/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Esclerosis Múltiple/metabolismo , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Western Blotting , Encéfalo/metabolismo , Encéfalo/patología , Movimiento Celular/fisiología , Células Cultivadas , Citometría de Flujo , Humanos , Inmunohistoquímica , Leucocitos Mononucleares/metabolismo , Esclerosis Múltiple/patologíaRESUMEN
Dense astrocytic scarring in chronic multiple sclerosis (MS) plaques produces an inhibitory environment which can impede tissue repair. Animal studies have shown that the antigenic phenotype of the most abundant cell type in the brain, the astrocyte, varies depending on astrocyte type and location. To identify the phenotype of scar astrocytes (SAs) in chronic lesions, markers of reactive astrocytes characterized in animal studies were investigated. To date these are the only established markers. Cerebral subventricular deep white matter from normal control, MS normal appearing white matter and lesions (acute, subacute and chronic) were examined by immunohistochemistry and immunoblotting. The antigenic profile of SAs revealed significant modification of astrocyte protein expression in chronic MS lesions. SAs express nestin, embryonic neural cell adhesion molecule, fibroblast growth factor receptor 4, epidermal growth factor receptor, nerve growth factor and a subpopulation of SAs also express basic fibroblast growth factor. These are in addition to the expected markers glial fibrillary acidic protein, vimentin, and the tenascins C and R. Therefore, an SA antigenic phenotype has now been defined. This knowledge may allow the development of therapeutic strategies that prevent scar formation and promote tissue repair.
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Astrocitos/fisiología , Esclerosis Múltiple/genética , Adulto , Anciano , Animales , Anticuerpos Monoclonales , Western Blotting , Corteza Cerebral/metabolismo , Corteza Cerebral/patología , Cicatriz , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Esclerosis Múltiple/patología , FenotipoRESUMEN
Disease progression in multiple sclerosis occurs within the interface of glial activation and gliosis. This study aimed to investigate the relationship between biomarkers of different glial cell responses: (i) to disease dynamics and the clinical subtypes of multiple sclerosis; (ii) to disability; and (iii) to cross-validate these findings in a post-mortem study. To address the first goal, 51 patients with multiple sclerosis [20 relapsing remitting (RR), 21 secondary progressive (SP) and 10 primary progressive (PP)] and 51 neurological control patients were included. Disability was assessed using the ambulation index (AI), the Expanded Disability Status Scale score (EDSS) and the 9-hole PEG test (9HPT). Patients underwent lumbar puncture within 7 days of clinical assessment. Post-mortem brain tissue (12 multiple sclerosis and eight control patients) was classified histologically and adjacent sites were homogenized for protein analysis. S100B, ferritin and glial-fibrillary acidic protein (GFAP) were quantified in CSF and brain-tissue homogenate by ELISA (enzyme-linked immunosorbent assay) techniques developed in-house. There was a significant trend for increasing S100B levels from PP to SP to RR multiple sclerosis (P < 0.05). S100B was significantly higher in RR multiple sclerosis than in control patients (P < 0.01), whilst ferritin levels were significantly higher in SP multiple sclerosis than in control patients (P < 0.01). The S100B : ferritin ratio discriminated patients with RR multiple sclerosis from SP, PP or control patients (P < 0.05, P < 0.01 and P < 0.01, respectively). Multiple sclerosis patients with poor ambulation (AI > or =7) or severe disability (EDSS >6.5) had significantly higher CSF GFAP levels than less disabled multiple sclerosis or control patients (P < 0.01 and P < 0.001, respectively). There was a correlation between GFAP levels and ambulation in SP multiple sclerosis (r = 0.57, P < 0.01), and between S100B level and the 9HPT in PP multiple sclerosis patients (r = -0.85, P < 0.01). The post-mortem study showed significantly higher S100B levels in the acute than in the subacute plaques (P < 0.01), whilst ferritin levels were elevated in all multiple sclerosis lesion stages. Both GFAP and S100B levels were significantly higher in the cortex of multiple sclerosis than in control brain homogenate (P < 0.001 and P < 0.05, respectively). We found that S100B is a good marker for the relapsing phase of the disease (confirmed by post-mortem observation) as opposed to ferritin, which is elevated throughout the entire course. GFAP correlated with disability scales and may therefore be a marker for irreversible damage. The results of this study have broad implications for finding new and sensitive outcome measures for treatment trials that aim to delay the development of disability. They may also be considered in future classifications of multiple sclerosis patients.
Asunto(s)
Esclerosis Múltiple/patología , Esclerosis Múltiple/fisiopatología , Neuroglía/patología , Proteínas S100 , Adulto , Anciano , Biomarcadores/análisis , Química Encefálica , Proteínas de Unión al Calcio/análisis , Proteínas de Unión al Calcio/líquido cefalorraquídeo , Líquido Cefalorraquídeo/química , Ensayo de Inmunoadsorción Enzimática , Femenino , Ferritinas/análisis , Ferritinas/líquido cefalorraquídeo , Proteína Ácida Fibrilar de la Glía/análisis , Proteína Ácida Fibrilar de la Glía/líquido cefalorraquídeo , Humanos , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/clasificación , Esclerosis Múltiple Crónica Progresiva/patología , Esclerosis Múltiple Crónica Progresiva/fisiopatología , Esclerosis Múltiple Recurrente-Remitente/patología , Esclerosis Múltiple Recurrente-Remitente/fisiopatología , Factores de Crecimiento Nervioso/análisis , Factores de Crecimiento Nervioso/líquido cefalorraquídeo , Neuroglía/metabolismo , Valor Predictivo de las Pruebas , Subunidad beta de la Proteína de Unión al Calcio S100 , Índice de Severidad de la EnfermedadRESUMEN
An increased level of myelin basic protein (MBP) degradation peptide 80-89, representative of myelin breakdown, is detected in myelinating foetal rat brain aggregate cultures supplemented with peritoneal macrophages at a time coinciding with the onset of myelination. During the period of myelination, the proportion of activated macrophages/microglia in the aggregates decreases, accompanied by a reduction in the content of MBP degradation products. During the recovery period following a demyelinating episode, the rate of MBP synthesis in antibody-treated standard aggregates was greater than in their medium controls. However, the rate of MBP accumulation was not as efficient in macrophage-enriched aggregates and was associated with persistently raised MBP peptide levels. Thus, as occurs in multiple sclerosis lesions, attempts at remyelination appear to be counterbalanced by macrophage-mediated demyelination, with the continued presence of degraded myelin rendering a local environment that is not fully conducive to remyelination.
Asunto(s)
Antígenos CD , Antígenos de Neoplasias , Antígenos de Superficie , Proteínas Aviares , Proteínas Sanguíneas , Encéfalo/embriología , Macrófagos/metabolismo , Esclerosis Múltiple/metabolismo , Proteína Básica de Mielina/metabolismo , Vaina de Mielina/metabolismo , Regeneración Nerviosa/inmunología , Fagocitosis/inmunología , Adulto , Animales , Anticuerpos/farmacología , Basigina , Encéfalo/inmunología , Encéfalo/metabolismo , Recuento de Células , Tamaño de la Célula/inmunología , Células Cultivadas/efectos de los fármacos , Células Cultivadas/inmunología , Células Cultivadas/metabolismo , Ectodisplasinas , Feto , Humanos , Inmunohistoquímica , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Glicoproteínas de Membrana/metabolismo , Proteínas de la Membrana/metabolismo , Persona de Mediana Edad , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/fisiopatología , Proteína Básica de Mielina/inmunología , Proteínas de la Mielina , Vaina de Mielina/inmunología , Vaina de Mielina/patología , Glicoproteína Asociada a Mielina/antagonistas & inhibidores , Glicoproteína Asociada a Mielina/inmunología , Glicoproteína Asociada a Mielina/metabolismo , Glicoproteína Mielina-Oligodendrócito , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Components of the plasminogen activator (PA) and matrix metalloprotease (MMP) cascade have been characterized in multiple sclerosis lesions by immunohistochemistry, enzyme-linked immunosorbent assay and enzyme activity assays in order to establish a functional role for the enzyme sequence in lesion development. Highly significant quantitative increases in urokinase PA (uPA), urokinase receptor (uPAR) and plasminogen activator inhibitor-1 were detected in acute multiple sclerosis lesions (P < 0.0001) and in uPAR in normal-appearing white matter (P < 0.0001) compared with control tissue. All three proteins were immunolocalized to mononuclear cells in perivascular cuffs and to macrophages in the lesion parenchyma. MMP-9 and the tissue inhibitor of metalloprotease-1 also increased during lesion development but the enzyme was present largely in the inactive pro-form. In contrast to uPA, the concentration and activity of tissue PA (tPA), the most abundant plasminogen activator in normal control brain, were reduced in multiple sclerosis specimens. In acute lesions tPA co-localized with fibrin(ogen) on large diameter axons also stained with SMI-32, an immunohistochemical marker of axonal damage. The uPA-uPAR complex, concentrated on inflammatory cells in the perivascular zone of the evolving lesion, may facilitate cellular infiltration into the CNS which is amplified by MMP- mediated degradation of blood vessel matrix. tPA localization on injured axons may be a marker of axonal damage or represent a protective mechanism aimed at removal of fibrin deposits and restoration of axonal function.
Asunto(s)
Axones/enzimología , Axones/patología , Esclerosis Múltiple/enzimología , Esclerosis Múltiple/patología , Activadores Plasminogénicos/metabolismo , Adulto , Anciano , Femenino , Humanos , Inflamación/enzimología , Inflamación/patología , Masculino , Metaloproteinasas de la Matriz/metabolismo , Persona de Mediana Edad , Inhibidor 1 de Activador Plasminogénico/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores del Activador de Plasminógeno Tipo Uroquinasa , Activador de Tejido Plasminógeno/metabolismo , Activador de Plasminógeno de Tipo Uroquinasa/metabolismoRESUMEN
Lewis rats, on recovery from monophasic clinical experimental allergic encephalomyelitis (EAE), can be induced to develop repeated paralytic relapses with a graded reduction in clinical severity following intraperitoneal administration of IL-12. By the time of the third relapse, the number and size of inflammatory cuffs in the spinal cord were reduced with the makeup of the cellular infiltrate shifting to a significantly increased number of B cells. Serum levels of myelin basic protein (MBP)-specific IgG1 and IgG2b were found to rise over time while MBP and MBP peptide-positive macrophages and microglia became evident in perivascular cuffs and in spinal cord parenchyma, indicative of myelin phagocytosis. Axonal death was observed in semithin and EM sections of spinal cord in third relapse animals in association with iNOS and tPA immunostaining throughout gray and white matter. These neurotoxic or excitotoxic agents may contribute to axonal damage directly or indirectly by activated microglia and macrophages, leading to limited damage of the axonal-myelin unit.
Asunto(s)
Axones/patología , Encefalomielitis Autoinmune Experimental/patología , Vaina de Mielina/patología , Enfermedad Aguda , Animales , Autoanticuerpos/biosíntesis , Axones/química , Encefalomielitis Autoinmune Experimental/inducido químicamente , Encefalomielitis Autoinmune Experimental/metabolismo , Femenino , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/química , Interleucina-12 , Interleucinas/sangre , Recuento de Leucocitos , Activación de Macrófagos , Proteína Básica de Mielina/inmunología , Parálisis/inducido químicamente , Ratas , Ratas Endogámicas Lew , Recurrencia , Médula Espinal/patología , Activador de Tejido Plasminógeno/análisis , Factor de Crecimiento Transformador beta/sangre , Factor de Crecimiento Transformador beta1RESUMEN
Insulin-like growth factors (IGFs) play an important role in development and myelination in the central nervous system (CNS) as well as in the proliferation and differentiation of cells of the immune system. To assess the influence of this growth factor family on demyelination and repair in multiple sclerosis (MS), the expression of IGF-I, IGF-II, insulin, IGF binding proteins (IGFBP) 1-3 and IGF-I receptor (IGF-IR) in CNS tissue from MS and normal control cases was studied by immunocytochemistry. In active MS lesions, the expression of IGF-I, insulin and IGFBP1 was detected in hypertrophic astrocytes while that of IGF-II and IGFBP2 and 3 was confined to foamy macrophages within lesions and activated microglia in adjacent white matter. IGF-IR, the major IGF receptor, was immunolocalized in macrophages and an astrocyte subpopulation in plaques. Oligodendrocytes in normal-appearing white matter expressed only IGFBP1, not IGFs or IGF-IR. As the remyelinating capacity of oligodendrocytes could be impaired owing to the absence of IGF-IR, the prevailing role of IGFs in inflammatory demyelination may be to promote phagocytosis of myelin and astrogliosis.
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Sistema Nervioso Central/metabolismo , Sistema Nervioso Central/patología , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Esclerosis Múltiple/metabolismo , Esclerosis Múltiple/patología , Somatomedinas/metabolismo , Adulto , Anciano , Enfermedades Desmielinizantes/metabolismo , Enfermedades Desmielinizantes/patología , Femenino , Humanos , Inmunohistoquímica , Macrófagos/metabolismo , Macrófagos/patología , Macrófagos/ultraestructura , Masculino , Persona de Mediana Edad , Oligodendroglía/metabolismo , Oligodendroglía/patología , Oligodendroglía/ultraestructura , FagocitosisRESUMEN
NF-kappaB is a transcription factor family which on translocation to the nucleus regulates gene expression during cell activation. As such, NF-kappaB may play a role in the microglial response to myelin damage in multiple sclerosis (MS) lesions. Here the cellular localization of NF-kappaB and expression of the inhibitory I kappaBalpha were examined by immunocytochemistry on central nervous system (CNS) tissue from MS and control cases. In normal control white matter, the active form of the NF-kappaB subunit RelA (p65) was localized in microglial nuclei, while the c-Rel and p50 subunits and the inhibitory I kappaBalpha were restricted to the cytoplasm. In contrast, in actively demyelinating plaques, the RelA, c-Rel, and p50 subunits of NF-kappaB and I kappaBalpha were all present in macrophage nuclei in both parenchymal and perivascular areas. RelA was also found in the nuclei of a subset of hypertrophic astrocytes. Only c-Rel had a nuclear localization in lymphocytes in perivascular inflammatory cuffs. Our results suggest that constitutive activation of the RelA subunit in the nuclei of resting microglia may facilitate a rapid response to pathological stimuli in the CNS. Activation of the inducible NF-kappaB pool in macrophages in MS lesions could amplify the inflammatory reaction through upregulation of NF-kappaB-controlled adhesion molecules and cytokines.
Asunto(s)
Encéfalo/patología , Proteínas de Unión al ADN/análisis , Proteínas I-kappa B , Macrófagos/patología , Microglía/patología , Esclerosis Múltiple/patología , FN-kappa B/análisis , Médula Espinal/patología , Adulto , Anciano , Encéfalo/citología , Encéfalo/inmunología , Neoplasias Encefálicas/patología , Núcleo Celular/patología , Núcleo Celular/ultraestructura , Infarto Cerebral/patología , Enfermedades Desmielinizantes/patología , Humanos , Inmunohistoquímica , Inflamación , Microglía/citología , Persona de Mediana Edad , Esclerosis Múltiple/inmunología , Inhibidor NF-kappaB alfa , FN-kappa B/antagonistas & inhibidores , Sustancia Gris Periacueductal/citología , Sustancia Gris Periacueductal/inmunología , Sustancia Gris Periacueductal/patología , Valores de Referencia , Médula Espinal/citologíaRESUMEN
The expression of tissue-type plasminogen activator (t-PA) and a number of metalloproteases as well as plasminogen activator inhibitor-1 (PAI-1) and tissue inhibitor of metalloproteases-1 (TIMP-1) was analyzed in the central nervous system (CNS) of normal control and multiple sclerosis (MS) cases by immunohistopathology. The expression of t-PA was detectable only in the blood vessel matrix in control white matter, but positive infiltrating mononuclear cells were also observed in MS white matter and primary lesions. In active plaques this pattern converted to strong positivity of foamy macrophages in areas of demyelination, declining in chronic lesions. In general PAI-1 expression paralleled that of t-PA. Gelatinase A and B were detected predominantly in astrocytes and microglia throughout normal control white matter, with additional positive mononuclear cells in perivascular cuffs in MS white matter. In the demyelinating lesion there is widespread prominent expression of gelatinase B in reactive astrocytes and macrophages, which persists in astrocytes in the chronic lesion. TIMP-1 was also present in the vessel matrix and in lesional macrophages. These observations on the coexpression of enzymes and inhibitors of the matrix degrading cascade in CNS tissue pinpoint t-PA, a rate-limiting enzyme, and gelatinase B as therapeutic targets in MS.
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Sistema Nervioso Central/metabolismo , Glicoproteínas/biosíntesis , Metaloendopeptidasas/biosíntesis , Esclerosis Múltiple/metabolismo , Proteínas de la Mielina/metabolismo , Proteínas del Tejido Nervioso/biosíntesis , Inhibidor 1 de Activador Plasminogénico/biosíntesis , Activador de Tejido Plasminógeno/biosíntesis , Adulto , Anciano , Sistema Nervioso Central/patología , Progresión de la Enfermedad , Inducción Enzimática , Femenino , Expresión Génica , Glicoproteínas/genética , Humanos , Hibridación in Situ , Masculino , Metaloendopeptidasas/genética , Persona de Mediana Edad , Esclerosis Múltiple/patología , Proteínas del Tejido Nervioso/genética , Inhibidor 1 de Activador Plasminogénico/genética , ARN Mensajero/análisis , Inhibidores Tisulares de Metaloproteinasas , Activador de Tejido Plasminógeno/genéticaRESUMEN
The work of the Pozega Medical Center Surgical Unit during the two war years (from July 16, 1991, to October 1, 1993) in Croatia is described. Since the war operations did not affect the town directly, the work was mostly characterized by treatment of wounds inflicted by explosive devices (51.2%) and localized mostly on the extremities (62.9%). Apart from these wounds, there were 0.7% burns, 26.6% gunshot wounds, and 19.6% injuries caused by blunt and hard objects. Of the total of 959 treated wounded persons, only 18 (1.9%) died, illustrating the success of the therapy administered according to the war surgery doctrine criteria.