RESUMO
Femtosecond pulse laser processing concentrates a huge quantity of light energy in extremely short pulses of a few tens to hundreds of femtoseconds, enabling superficial laser machining or marking of any kind of materials, with a reduced or insignificant heat affected area. A digitized paper printed image of the face on the Turin Shroud was used to monitor a scan head intercalated between a femtosecond pulsed laser source and a linen fabric sample, enabling the direct 2D reproduction of the image of the face with a laser beam size corresponding to one pixel of the digitized image. The contrast in the marked image was controlled by adjusting the energy density, the number of superimposed pulses per pixel, and the distance between successive impacts. The visual aspect of the laser-induced image is very similar, at naked eye, to the source image. The negative photograph of the marked linen fabric reveals a face remarkably close to the well-known negative picture of the face on the Turin Shroud. Analyses by infrared spectroscopy, Raman spectroscopy, and scanning electron microscopy were performed to characterize the laser marked areas.
RESUMO
The neuroprotective effects of a systemically active, highly selective, corticotropin-releasing factor-1 (CRF1) receptor antagonist, R121920 ((7-(dipropylamino)-2,5-dimethyl-3- [2-(dimethylamino)-5-pyridyl] pyrazolo [1,5-a] pyrimidine), was assessed in two rat models of permanent focal cerebral ischemia, where the middle cerebral artery (MCA) was occluded either through the subtemporal approach or using the intraluminal suture technique. R121920 rapidly crossed the blood-brain barrier after intravenous (IV) bolus administration (10 mg/kg), with peak brain concentrations at 5 minutes (2.26 +/- 0.40 microg/mL), which were approximately 2-fold greater than those in plasma (0.98 +/- 0.24 microg/mL). Treatment with R121920 (10 mg/kg IV followed by 5 mg/kg subcutaneously at hourly intervals for 4 hours) significantly (P < 0.001) reduced total (by 40%) and cortical (by 37%) infarct volume at 24 hours after subtemporal MCA occlusion (MCAO). In the intraluminal suture MCAO model, IV administration of R121920 (10 mg/kg) at the time of ischemia onset (and at multiple times thereafter) reduced both hemispheric infarct volume (by 34%, P < 0.001) and brain swelling (by 50%, P < 0.001) when assessed at 24 hours. In this model of focal ischemia, significant reduction (P < 0.05) in both outcome measures was obtained when R121920 administration was delayed up to 1 hour after MCAO. These results further define the antiischemic properties of selective CRF 1 antagonists in two experimental models of permanent focal cerebral ischemia.
Assuntos
Isquemia Encefálica/fisiopatologia , Fármacos Neuroprotetores/farmacologia , Receptores de Hormônio Liberador da Corticotropina/antagonistas & inibidores , Animais , Edema Encefálico/fisiopatologia , Edema Encefálico/prevenção & controle , Masculino , Artéria Cerebral Média/patologia , Ratos , Ratos Sprague-DawleyRESUMO
Nitric oxide (NO), produced by inducible NO synthase (iNOS), may play a role in inflammatory demyelinating diseases of the central nervous system (CNS). We show upregulation of iNOS mRNA in CNS of SJL/J mice with experimental allergic encephalomyelitis (EAE). Using antibodies against mouse iNOS, GFAP (a marker for astrocytes) and Mac-1/CD11b (a marker for macrophages/microglia), both astrocytes and macrophages/microglia were identified as iNOS-expressing cells in situ in EAE lesions. GFAP + astrocytes not associated with inflammatory infiltrates were also found to express iNOS. Because microglia rather than astrocytes are implicated in demyelinating pathology, we propose that microglial NO may be cytopathic whereas astrocyte-derived NO may be protective in EAE.
Assuntos
Astrócitos/enzimologia , Encefalomielite Autoimune Experimental/enzimologia , Microglia/enzimologia , Óxido Nítrico Sintase/metabolismo , Animais , Astrócitos/metabolismo , Sistema Nervoso Central/citologia , Sistema Nervoso Central/enzimologia , Sistema Nervoso Central/fisiopatologia , Encefalomielite Autoimune Experimental/genética , Indução Enzimática , Feminino , Expressão Gênica , Proteína Glial Fibrilar Ácida/metabolismo , Camundongos , Camundongos Endogâmicos , Óxido Nítrico Sintase/genética , Distribuição TecidualRESUMO
Insulin-like growth factors (IGFs) are important trophic factors during development as well as in the adult or damaged nervous system. Their trophic actions are modulated by interactions with six distinct IGF binding proteins. The mRNA expression profiles of binding proteins 2, 4 and 5 in the normal developing and adult CNS are well characterized and are shown to have distinctive, non-overlapping distributions. The IGF binding protein-6 (BP6) is also expressed in the CNS, however, details regarding its mRNA expression distribution in the developing and adult nervous system is limited. BP6 has the unique property of preferentially binding the IGF-II ligand. Coupled with the fact that this ligand is the most abundantly expressed IGF in the adult CNS, this suggests that the IGF-II/BP6 complex has a unique role in modulating IGF-II function in the adult brain. In this report the anatomical distribution of BP6 messenger RNA in the developing and adult rat nervous system is presented. In the embryonic animal the CNS expression is tightly restricted to trigeminal ganglia and, relative to the rest of the embryo, this structure has the highest expression. The expression in the forebrain and cerebellum does not occur until after postnatal day 21 and then is primarily associated with GABAergic interneurons. The highest levels of expression in the adult animal are in the hindbrain, spinal cord, cranial ganglia, and dorsal root ganglia. These nuclei in the hindbrain and periphery that express BP6 are all associated with the coordination of sensorimotor function in the cerebellum, which indicates an important role for the BP6/IGF-II complex in the function and maintenance of these systems.
Assuntos
Encéfalo/metabolismo , Gânglios Sensitivos/metabolismo , Proteína 6 de Ligação a Fator de Crescimento Semelhante à Insulina/biossíntese , Sistema Nervoso/metabolismo , Medula Espinal/metabolismo , Animais , Imuno-Histoquímica , Técnicas In Vitro , Fator de Crescimento Insulin-Like I/metabolismo , Fator de Crescimento Insulin-Like II/metabolismo , RNA Mensageiro/biossíntese , Ratos , Distribuição Tecidual , Regulação para CimaRESUMO
The neuronal apoptosis inhibitory protein (NAIP) is known to have anti-apoptotic functions, and its gene is often mutated in severe cases of spinal muscular atrophy (SMA), a disease characterized by motor neuron degeneration. In this study, we examined the distribution of the endogenous NAIP protein in normal human spinal cord and brain tissue by using a polyclonal antibody against NAIP. Immunohistochemical staining demonstrated that NAIP is strongly expressed in anterior horn and motor cortex neurons of normal brains, and it is not altered in the remaining motor neurons of patients with amyotrophic lateral sclerosis (ALS). NAIP is also located in human fetal neurons and in adult choroid plexus cells. These results suggest that the anti apoptotic molecule NAIP may be important in motor neurons, but it specifically does not appear to be altered in ALS.
Assuntos
Apoptose/fisiologia , Química Encefálica/fisiologia , Encéfalo/anatomia & histologia , Proteínas do Tecido Nervoso/metabolismo , Medula Espinal/anatomia & histologia , Medula Espinal/metabolismo , Adenoviridae/genética , Adulto , Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/metabolismo , Esclerose Lateral Amiotrófica/patologia , Células do Corno Anterior/metabolismo , Células do Corno Anterior/patologia , Encéfalo/patologia , Química Encefálica/genética , Células Cultivadas , Células HeLa , Humanos , Imuno-Histoquímica , Microscopia Confocal , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Proteínas do Tecido Nervoso/genética , Proteína Inibidora de Apoptose Neuronal , Neurônios/metabolismo , Inclusão em Parafina , Medula Espinal/patologia , Transdução Genética/genéticaRESUMO
CC chemokine receptor 7 (CCR7) is involved in the initiation of immune responses by mediating the migration of naïve T cells and mature dendritic cells to T-cell-rich zones of secondary lymphoid organs where antigen presentation occurs. To address whether CCR7 plays a role in the development of autoimmunity, we induced experimental autoimmune encephalomyelitis in CCR7-deficient mice on a C57BL/6 background (CCR7(-/-)) using the neuroantigen, myelin oligodendrocyte glycoprotein 35-55 amino acid peptide (MOG((35-55))) and Bordetella pertussis toxin (PTX). CCR7(-/-) mice acquired disease with an intensity similar to wild-type littermates. MOG((35-55))-specific lymphocyte responses were dominant in the spleen of CCR7(-/-) mice, rather than in lymph nodes as observed in wild-type mice. These results indicate that effective immune responses (with altered kinetics) can develop in the absence of CCR7 but develop in the spleen rather than lymph nodes as CCR7 is necessary for T and dendritic cells to enter lymph nodes.
Assuntos
Encefalomielite Autoimune Experimental/genética , Encefalomielite Autoimune Experimental/imunologia , Receptores de Quimiocinas/deficiência , Receptores de Quimiocinas/genética , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Sequência de Aminoácidos , Animais , Movimento Celular/genética , Movimento Celular/imunologia , Células Cultivadas , Quimiocinas CC/metabolismo , Doença Crônica , Células Dendríticas/citologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Encefalomielite Autoimune Experimental/patologia , Feminino , Imunofenotipagem , Linfonodos/citologia , Linfonodos/imunologia , Linfonodos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dados de Sequência Molecular , Receptores CCR7 , Receptores de Quimiocinas/fisiologia , Baço/imunologia , Baço/metabolismo , Baço/patologia , Subpopulações de Linfócitos T/patologiaRESUMO
Activated glial cells are implicated in regulating and effecting the immune response that occurs within the CNS as part of multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). The peripheral benzodiazepine receptor (PBR) is expressed in glial cells. We examined the utility of using in vitro and in vivo ligand binding to the PBR as a measure of lesion activity in autoimmune CNS demyelinating diseases. Applying a combined autoradiography and immunohistochemical approach to spinal cord and brain tissues from mice with EAE, we found a correlation at sites of inflammatory lesions between [3H]-PK11195 binding and immunoreactivity for the activated microglial/macrophage marker Mac-1/CD11b. In MS tissues, [3H]-PK11195 binding correlated with sites of immunoreactivity for the microglial/macrophage marker CD68, at the edges of chronic active plaques. Positron emission tomography (PET) imaging with [11C]-PK11195 showed ligand uptake only at sites of active MS lesions defined by magnetic resonance imaging criteria. Our results indicate the potential to develop markers suitable for both in vitro and in vivo use, which will serve to help correlate phenotypic and functional properties of cells which participate in disease or injury responses within the CNS.
Assuntos
Encefalomielite Autoimune Experimental/metabolismo , Encefalomielite Autoimune Experimental/fisiopatologia , Isoquinolinas/metabolismo , Microglia/fisiologia , Esclerose Múltipla/metabolismo , Esclerose Múltipla/fisiopatologia , Receptores de GABA-A/metabolismo , Animais , Biomarcadores , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Encéfalo/patologia , Encefalomielite Autoimune Experimental/patologia , Humanos , Imageamento por Ressonância Magnética , Meninges/metabolismo , Camundongos , Camundongos Endogâmicos , Esclerose Múltipla/patologia , Valores de Referência , Tomografia Computadorizada de EmissãoRESUMO
BACKGROUND AND OBJECTIVE: Meropenem is the first of a new class of carbapenems which may be administered without cilastatin. This study was performed to assess the clinical efficacy and tolerability of meropenem monotherapy (1 g/8 h) compared with the standard combination of ceftazidime (2 g/8 h) plus amikacin (15 mg/kg/day) for the empirical treatment of infective febrile episodes in neutropenic cancer patients. METHODS: This was a three-center, randomized, non-blind parallel group trial. The primary objective was to compare the clinical efficacy of meropenem monotherapy with that of ceftazidime plus amikacin in the empirical treatment of febrile infective episodes in neutropenic patients. This was evaluated by the number of patients surviving on unmodified therapy at 72 h (primary end point) and by the clinical response at the end of therapy (secondary end point). RESULTS: A total of 93 febrile episodes (46 meropenem, 47 ceftazidime/amikacin) were evaluable. Bone marrow transplant patients accounted for 49.5% of all cases. There was a high incidence of Gram-positive infections but no pseudomonal infections. Microbiologically documented infections, clinically documented infections and unexplained fever accounted for 45%, 10% and 45% of episodes, respectively. There was a similar proportion of patients in the meropenem and ceftazidime/amikacin groups on unmodified empiric therapy at 72 h (80.4% vs 76.6%, p = 0.65,) and cured at the end of therapy (37% vs 36.2%, p = 0.9). No significant difference in tolerability was observed between the groups. Meropenem was well tolerated; of note, there were no cases of nausea/vomiting or seizure related to its use. INTERPRETATION AND CONCLUSIONS: Meropenem monotherapy was well tolerated and produced response rates similar to those obtained with ceftazidime/amikacin. The low overall success rates with both treatments concur with those of other recent studies and are probably due to a combination of several factors, including the adoption of strict assessment criteria.
Assuntos
Quimioterapia Combinada/uso terapêutico , Febre/tratamento farmacológico , Neutropenia/tratamento farmacológico , Amicacina/uso terapêutico , Ceftazidima/uso terapêutico , Febre/etiologia , Humanos , Meropeném , Neoplasias/complicações , Neutropenia/complicações , Tienamicinas/uso terapêuticoRESUMO
The cytokine interferon-gamma (IFNgamma) is implicated in the induction of acute CNS inflammation, but it is less clear what role if any IFNgamma plays in progression to chronic demyelination and neurological deficit. To address this issue, we have expressed IFNgamma in myelinating oligodendrocytes of transgenic mice. MHC I immunostaining and iNOS mRNA were upregulated in their CNS, but such transgenic mice showed no spontaneous CNS inflammation or demyelination, and the incidence, severity, and histopathology of experimental autoimmune encephalomyelitis (EAE) were similar to nontransgenic controls. In contrast to control mice, which remit from EAE with resolution of glial reactivity and leukocytic infiltration, transgenics showed chronic neurological deficits. While activated microglia/macrophages persisted in demyelinating lesions for over 100 days, CD4(+) T lymphocytes were no longer present in CNS. IFNgamma therefore may play a role in chronic demyelination and long-term disability following the induction of demyelinating disease. Because IFNgamma may have neural as well as immune-infiltrating origins, these findings generate a new perspective on its role in the CNS.
Assuntos
Doenças Desmielinizantes/imunologia , Encefalomielite Autoimune Experimental/imunologia , Interferon gama/genética , Interferon gama/imunologia , Animais , Complexo CD3/análise , Linfócitos T CD4-Positivos/química , Linfócitos T CD4-Positivos/imunologia , Doença Crônica , Progressão da Doença , Feminino , Expressão Gênica/imunologia , Antígenos Comuns de Leucócito/análise , Antígeno de Macrófago 1/análise , Macrófagos/química , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microglia/química , Microglia/imunologia , Gravidez , RNA Mensageiro/análiseRESUMO
Recent evidence suggests that spinal cord glia can contribute to enhanced nociceptive responses. However, the signals that cause glial activation are unknown. Fractalkine (CX3C ligand-1; CX3CL1) is a unique chemokine expressed on the extracellular surface of spinal neurons and spinal sensory afferents. In the dorsal spinal cord, fractalkine receptors are primarily expressed by microglia. As fractalkine can be released from neurons upon strong activation, it has previously been suggested to be a neuron-to-glial signal that induces glial activation. The present series of experiments provide an initial investigation of the spinal pain modulatory effects of fractalkine. Intrathecal fractalkine produced dose-dependent mechanical allodynia and thermal hyperalgesia. In addition, a single injection of fractalkine receptor antagonist (neutralizing antibody against rat CX3C receptor-1; CX3CR1) delayed the development of mechanical allodynia and/or thermal hyperalgesia in two neuropathic pain models: chronic constriction injury (CCI) and sciatic inflammatory neuropathy. Intriguingly, anti-CX3CR1 reduced nociceptive responses when administered 5-7 days after CCI, suggesting that prolonged release of fractalkine may contribute to the maintenance of neuropathic pain. Taken together, these initial investigations of spinal fractalkine effects suggest that exogenous and endogenous fractalkine are involved in spinal sensitization, including that induced by peripheral neuropathy.
Assuntos
Quimiocinas CX3C/metabolismo , Proteínas de Membrana/metabolismo , Neuroglia/metabolismo , Nociceptores/fisiologia , Dor/metabolismo , Doenças do Sistema Nervoso Periférico/metabolismo , Medula Espinal/metabolismo , Animais , Anticorpos/farmacologia , Receptor 1 de Quimiocina CX3C , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/fisiologia , Quimiocina CX3CL1 , Quimiocinas CX3C/administração & dosagem , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Hiperalgesia/induzido quimicamente , Hiperalgesia/metabolismo , Hiperalgesia/fisiopatologia , Injeções Espinhais , Ligadura , Masculino , Proteínas de Membrana/administração & dosagem , Proteínas de Membrana/efeitos dos fármacos , Microglia/efeitos dos fármacos , Microglia/metabolismo , Neuroglia/efeitos dos fármacos , Neurônios/metabolismo , Nociceptores/efeitos dos fármacos , Dor/induzido quimicamente , Dor/fisiopatologia , Doenças do Sistema Nervoso Periférico/induzido quimicamente , Doenças do Sistema Nervoso Periférico/fisiopatologia , Ratos , Ratos Sprague-Dawley , Receptores de Citocinas/antagonistas & inibidores , Receptores de Citocinas/metabolismo , Receptores de HIV/antagonistas & inibidores , Receptores de HIV/metabolismo , Neuropatia Ciática/induzido quimicamente , Neuropatia Ciática/metabolismo , Neuropatia Ciática/fisiopatologia , Medula Espinal/efeitos dos fármacos , Medula Espinal/fisiopatologiaRESUMO
Proteolytic processing of amyloid precursor protein (APP) through an endosomal/lysosomal pathway generates carboxy-terminal polypeptides that contain an intact beta-amyloid domain. Cleavage by as-yet unidentified proteases releases the beta-amyloid peptide in soluble form. In Alzheimer's disease, aggregated beta-amyloid is deposited in extracellular neuritic plaques. Although most of the molecular mechanisms involving beta-amyloid and APP in the aetiology of Alzheimer's disease are still unclear, changes in APP metabolism may be important in the pathogenesis of the disease. Here we show that transgenic mice expressing the amyloidogenic carboxy-terminal 104 amino acids of APP develop, with ageing, extracellular beta-amyloid immunoreactivity, increased gliosis and microglial reactivity, as well as cell loss in the CA1 region of the hippocampus. Adult transgenic mice demonstrate spatial-learning deficits in the Morris water maze and in maintenance of long-term potentiation (LTP). Our results indicate that alterations in the processing of APP may have considerable physiological effects on synaptic plasticity.