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α-Hydroxy ketones are a class of vital organic skeletons that generally exist in a variety of natural products and high-value chemicals. However, the traditional synthetic route for their production involves toxic Hg salts and corrosive H2SO4 as catalysts, resulting in harsh conditions and the undesired side reaction of Meyer-Schuster rearrangement. In this study, CO2-promoted hydration of propargylic alcohols was achieved for the synthesis of various α-hydroxy ketones. Notably, this process was catalyzed using an environmentally friendly and cost-effective biomass-based ionic liquids/CuCl system, which effectively eliminated the side reaction. The ionic liquids utilized in this system are derived from natural biomass materials, which exhibited recyclability and catalytic activity under 1 bar of CO2 pressure without volatile organic solvents or additives. Evaluation of the green metrics revealed the superiority of this CuCl/ionic liquid system in terms of environmental sustainability. Further mechanistic investigation attributed the excellent performance to the ionic liquid component, which exhibited multifunctionality in activating substrates, CO2 and the Cu component.
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Alcinos , Líquidos Iônicos , Propanóis , Cetonas , Dióxido de Carbono , Biomassa , CatáliseRESUMO
PURPOSE: Melanotic cells with large spherical melanosomes, thought to originate from retinal pigment epithelium (RPE), are found in eyes with neovascular age-related macular degeneration (nvAMD). To generate hypotheses about RPE participation in fibrosis, we correlate histology to clinical imaging in an eye with prominent black pigment in fibrotic scar secondary to nvAMD. METHODS: Macular findings in a white woman with untreated inactive subretinal fibrosis due to nvAMD in her right eye were documented over 9 years with color fundus photography (CFP), fundus autofluorescence (FAF) imaging, and optical coherence tomography (OCT). After death (age 90 years), this index eye was prepared for light and electron microscopy to analyze 7 discrete zones of pigmentation in the fibrotic scar. In additional donor eyes with nvAMD, we determined the frequency of black pigment (n = 36 eyes) and immuno-labeled for retinoid, immunologic, and microglial markers (RPE65, CD68, Iba1, TMEM119; n = 3 eyes). RESULTS: During follow-up of the index eye, black pigment appeared and expanded within a hypoautofluorescent fibrotic scar. The blackest areas correlated to melanotic cells (containing large spherical melanosomes), some in multiple layers. Pale areas had sparse pigmented cells. Gray areas correlated to cells with RPE organelles entombed in the scar and multinucleate cells containing sparse large spherical melanosomes. In 94% of nvAMD donor eyes, hyperpigmentation was visible. Certain melanotic cells expressed some RPE65 and mostly CD68. Iba1 and TMEM119 immunoreactivity, found both in retina and scar, did not co-localize with melanotic cells. CONCLUSION: Hyperpigmentation in CFP results from both organelle content and optical superimposition effects. Black fundus pigment in nvAMD is common and corresponds to cells containing numerous large spherical melanosomes and superimposition of cells containing sparse large melanosomes, respectively. Melanotic cells are molecularly distinct from RPE, consistent with a process of transdifferentiation. The subcellular source of spherical melanosomes remains to be determined. Detailed histology of nvAMD eyes will inform future studies using technologies for spatially resolved molecular discovery to generate new therapies for fibrosis. The potential of black pigment as a biomarker for fibrosis can be investigated in clinical multimodal imaging datasets.
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Neovascularização de Coroide/complicações , Hiperpigmentação/patologia , Melanossomas/ultraestrutura , Retina/patologia , Degeneração Macular Exsudativa/complicações , Idoso de 80 Anos ou mais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Feminino , Fibrose , Humanos , Hiperpigmentação/etiologia , Hiperpigmentação/metabolismo , Masculino , Melanossomas/metabolismo , Proteínas de Membrana/metabolismo , Proteínas dos Microfilamentos/metabolismo , Retina/metabolismo , Estudos Retrospectivos , Tomografia de Coerência Óptica , Acuidade Visual , cis-trans-Isomerases/metabolismoRESUMO
Understanding the molecular composition of ocular tissues and fluids could inform new approaches to prevalent causes of blindness. Subretinal fluid accumulating between the photoreceptor outer segments and retinal pigment epithelium (RPE) is potentially a rich source of proteins and lipids normally cycling among outer retinal cells and choroid. Herein, intact post-translationally modified proteins (proteoforms) were extracted from subretinal fluids of five patients with rhegmatogenous retinal detachment (RRD), analyzed by tandem mass spectrometry, and compared to published data on these same proteins as synthesized by other organs. Single-nuclei transcriptomic data from non-diseased human retina/RPE were used to identify whether proteins in subretinal fluid were of potential ocular origin. Two human donor eyes with normal maculas were immunoprobed for transthyretin (TTR) with appropriate controls. The three most abundant proteins detected in subretinal fluid were albumin, TTR, and apolipoprotein A-I. Remarkably, TTR relative to the other proteins was more abundant than its serum counterpart, suggestive of TTR being synthesized predominantly locally. Six proteoforms of TTR were detected, with the relative amount of glutathionylated TTR being much higher in the subretinal fluid (12-43%) than values reported for serum (<5%) and cerebrospinal fluid (0.4-13%). Moreover, a putative glycosylated TTR dimer of 32,428 Da was detected as the fourth most abundant protein. The high abundance of TTR and putative TTR dimer in subretinal fluid was supported by analysis of available single-nuclei transcriptomic data, which showed strong and specific signal for TTR in RPE. Immunohistochemistry further showed strong diffuse TTR immunoreactivity in choroidal stroma that contrasted with vertically aligned signal in the outer segment zone of the subretinal space and negligible signal in RPE cell bodies. These results suggest that TTR in the retina is synthesized intraocularly, and glutathionylation is crucial for its normal function. Further studies on the composition, function, and quantities of TTR and other proteoforms in subretinal fluid could inform mechanisms, diagnostic methods, and treatment strategies for age-related macular degeneration, familial amyloidosis, and other retinal diseases involving dysregulation of physiologic lipid transfer and oxidative stress.
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Descolamento Retiniano , Doenças Retinianas , Humanos , Pré-Albumina/genética , Descolamento Retiniano/metabolismo , Doenças Retinianas/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Líquido Sub-Retiniano/metabolismoRESUMO
Autophagy and lysosomal function are important for protein homeostasis and their dysfunction have been associated with Alzheimer's disease (AD). Increased immunoreactivities of an important lysosomal protease, cathepsin D (Cat D), are evident in amyloid plaques and neurons in patients with AD. This study tests the hypothesis that deleting one allele of the cathepsin D gene (Ctsd) impacts cerebral ß-amyloidosis in amyloid-ß precursor protein (APP)sw/PS1dE9 (APP/PS1) double transgenic mice. Despite a significant 38% decrease in Cat D level in APP/PS1/Ctsd+/- compared with APP/PS1/Ctsd+/+ mice, no changes in steady state levels and deposition of Aß were found in the brain. There were also no differences in APP processing, the levels of two other Aß-degrading proteases, the levels of autophagy related protein, such as LAMP2, P62, LC3-I, LC3-II, and Beclin-1, or the markers of neuroinflammation, observed between the APP/PS1/Ctsd+/+ and APP/PS1/Ctsd+/- mice. Our findings demonstrate that in wild-type mice, Cat D protein levels are either in excess or redundant with other factors in the brain, and at least one allele of Ctsd is dispensable for cerebral ß-amyloidosis and autophagy in APP/PS1 transgenic mice.
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Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/metabolismo , Precursor de Proteína beta-Amiloide/metabolismo , Autofagia , Encéfalo/metabolismo , Catepsina D/metabolismo , Doença de Alzheimer/genética , Precursor de Proteína beta-Amiloide/genética , Animais , Autofagia/genética , Autofagia/fisiologia , Catepsina D/genética , Modelos Animais de Doenças , Camundongos Transgênicos , Neurônios/metabolismo , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Placa Amiloide/metabolismoRESUMO
Isoprenoids and prenylated proteins have been implicated in the pathophysiology of Alzheimer disease (AD), including amyloid-ß precursor protein metabolism, Tau phosphorylation, synaptic plasticity, and neuroinflammation. However, little is known about the relative importance of the two protein prenyltransferases, farnesyltransferase (FT) and geranylgeranyltransferase-1 (GGT), in the pathogenesis of AD. In this study, we defined the impact of deleting one copy of FT or GGT on the development of amyloid-ß (Aß)-associated neuropathology and learning/memory impairments in APPPS1 double transgenic mice, a well established model of AD. Heterozygous deletion of FT reduced Aß deposition and neuroinflammation and rescued spatial learning and memory function in APPPS1 mice. Heterozygous deletion of GGT reduced the levels of Aß and neuroinflammation but had no impact on learning and memory. These results document that farnesylation and geranylgeranylation play differential roles in AD pathogenesis and suggest that specific inhibition of protein farnesylation could be a potential strategy for effectively treating AD.
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Doença de Alzheimer/genética , Doença de Alzheimer/fisiopatologia , Cognição , Farnesiltranstransferase/deficiência , Farnesiltranstransferase/genética , Deleção de Genes , Doença de Alzheimer/enzimologia , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/metabolismo , Animais , Encéfalo/metabolismo , Modelos Animais de Doenças , Memória , Camundongos , Camundongos Transgênicos , ProteóliseRESUMO
The post-heat mechanical property is one of the important indices for the fire-resistance evaluation of fiber-reinforced polymers. At present, the primary approach to improving the post-heat mechanical property of a material involves incorporating inorganic fillers; yet, the enhancement is limited, and is accompanied by a reduction in room-temperature performance and processability. This study prepares glass-fiber-reinforced composites with elevated mechanical properties after heat through utilizing two variants of epoxy resins modified with polysiloxane, phenolic resin, kaolin, and graphite. In comparison to the phenolic samples, the phenylpropylsiloxane-modified epoxy resulted in a 115% rise in post-heat flexural strength and a 70% increase in the room-temperature flexural strength of phenolic composites. On the other hand, dimethylsiloxane-modified epoxy leads to a 117% improvement in post-heat flexural strength but a 44% decrease in the room-temperature flexural strength of phenolic composites. Macroscopic/microscopic morphologies and a residual structure model of the composites after heat reveal that, during high temperature exposure, the pyrolysis products of polysiloxane promote interactions between carbon elements and fillers, thus preserving more residues and improving the dimensional stability as well as the density of materials. Consequently, a notable enhancement is observed in both the post-heat flexural strength and the mass of carbon residue after the incorporation of polysiloxane and fillers into the materials. The pyrolysis products of polysiloxane-modified epoxy play a vital role in enhancing the post-heat flexural strength by promoting carbon retention, carbon fixation, and interactions with fillers, offering novel pathways for the development of advanced composites with superior fire-resistance properties.
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The fovea of the human retina, a specialization for acute and color vision, features a high concentration of cone photoreceptors. A pit on the inner retinal aspect is created by the centrifugal migration of post-receptoral neurons. Foveal cells are specified early in fetal life, but the fovea reaches its final configuration postnatally. Pre-term birth retards migration resulting in a small pit, a small avascular zone, and nearly continuous inner retinal layers. To explore the involvement of Müller glia, we used serial-section electron microscopic reconstructions to examine the morphology and neural contacts of Müller glia contacting a single foveal cone in a 28-year-old male organ donor born at 28 weeks of gestation. A small non-descript foveal avascular zone contained massed glial processes that included a novel class of 'inner' Müller glia. Similar to classic 'outer' Müller glia that span the retina, inner Müller glia have bodies in the inner nuclear layer (INL). These cells are densely packed with intermediate filaments and insert processes between neurons. Unlike 'outer' Müller glia, 'inner' Müller glia do not reach the external limiting membrane but instead terminate at the outer plexiform layer. One completely reconstructed inner cell ensheathed cone pedicles and a cone-driven circuit of midget bipolar and ganglion cells. Inner Müller glia outnumber foveal cones by 1.8-fold in the outer nuclear layer (221,448 vs. 123,026 cells/mm2). Cell bodies of inner Müller glia outnumber those of outer Müller glia by 1.7-fold in the INL (41,872 vs. 24,631 cells/ mm2). Müller glia account for 95 and 80% of the volume of the foveal floor and Henle fiber layer, respectively. Determining whether inner cells are anomalies solely resulting from retarded lateral migration of inner retinal neurons in pre-term birth requires further research.
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This paper presents an easy and low-cost flame treatment method to improve the bonding performance of GF/EP (Glass Fiber-Reinforced Epoxy) pultrusion plates, which are using widely for large size wind blades. In order to explore the effect of flame treatment on the bonding performance of the precast GF/EP pultruded sheet vs. the infusion plate, the GF/EP pultruded sheets were treated with different flame treatment cycles and were embedded in the fiber fabrics during the vacuum-assisted resin infusion process (VARI). The bonding shear strengths were measured by tensile shear tests. It is found that after 1, 3, 5, and 7 flame treatments, the tensile shear strength between the GF/EP pultrusion plate and infusion plate increased by 8.0%, 13.3%, 22.44%, and -2.1%, respectively. This indicates that the maximum tensile shear strength can be obtained after five times of flame treatment. In addition, DCB and ENF tests were also adopted to characterize the fracture toughness of the bonding interface with the optimal flame treatment. It is found that the optimal treatment gives increments of 21.84% and 78.36% for G I C and G II C, respectively. Finally, the surficial topography of the flame-treated GF/EP pultruded sheets were characterized by optical microscopy, SEM, contact angle test, FTIR, and XPS. The results show that flame treatment plays an impact on the interfacial performance through the combination of physical meshing locking and chemical bonding mechanism. Proper flame treatment would remove the weak boundary layer and mold release agent on the surface of the GF/EP pultruded sheet, etch the bonding surface and improve the oxygen-containing polar groups, such as C-O and O-C=O, to improve the surface roughness and surface tension coefficient of pultruded sheet to enhance the bonding performance. Excessive flame treatment destroys the integrity of epoxy matrix on bonding surface which results into the exposure of the glass fiber, and the carbonization of release agent and resin on the surface loosen the surficial structure, which reduces the bonding properties.
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In filament wound composites, fiber bundles cross each other and form an undulating architecture, which may significantly affect the mechanical behavior of composites. In this study, the tensile mechanical behavior of filament wound laminates was studied experimentally and numerically, and the influences of the bundle thickness and winding angle on the mechanical behavior of the filament wound plates were also explored. In the experiments, tensile tests were carried out on filament wound plates and laminated plates. It was found that, compared to laminated plates, filament wound plates had lower stiffness, greater failure displacement, similar failure loads, and more obvious strain concentration areas. In numerical analysis, mesoscale finite element models, which take into account the fiber bundles' undulating morphology, were created. The numerical predictions correlated well with the experimental ones. Further numerical studies have shown that the stiffness reduction coefficient of filament wound plates with a winding angle of ±55° decreased from 0.78 to 0.74 as the bundle thickness increased from 0.4 mm to 0.8 mm. The stiffness reduction coefficients of filament wound plates with wound angles of ±15°, ±25°, and ±45° were 0.86, 0.83, and 0.8, respectively.
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Mitochondria are candidate reflectivity signal sources in optical coherence tomography (OCT) retinal imaging. Here, we use deep-learning-assisted volume electron microscopy of human retina and in vivo imaging to map mitochondria networks in the outer plexiform layer (OPL), where photoreceptors synapse with second-order interneurons. We observed alternating layers of high and low mitochondrial abundance in the anatomical OPL and adjacent inner nuclear layer (INL). Subcellular resolution OCT imaging of human eyes revealed multiple reflective bands that matched the corresponding INL and combined OPL sublayers. Data linking specific mitochondria to defined bands in OCT may help improve clinical diagnosis and the evaluation of mitochondria-targeting therapies.
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Introduction: Age related macular degeneration (AMD) causes legal blindness worldwide, with few therapeutic targets in early disease and no treatments for 80% of cases. Extracellular deposits, including drusen and subretinal drusenoid deposits (SDD; also called reticular pseudodrusen), disrupt cone and rod photoreceptor functions and strongly confer risk for advanced disease. Due to the differential cholesterol composition of drusen and SDD, lipid transfer and cycling between photoreceptors and support cells are candidate dysregulated pathways leading to deposit formation. The current study explores this hypothesis through a comprehensive lipid compositional analysis of SDD. Methods: Histology and transmission electron microscopy were used to characterize the morphology of SDD. Highly sensitive tools of imaging mass spectrometry (IMS) and nano liquid chromatography tandem mass spectrometry (nLC-MS/MS) in positive and negative ion modes were used to spatially map and identify SDD lipids, respectively. An interpretable supervised machine learning approach was utilized to compare the lipid composition of SDD to regions of uninvolved retina across 1873 IMS features and to automatically discern candidate markers for SDD. Immunohistochemistry (IHC) was used to localize secretory phospholipase A2 group 5 (PLA2G5). Results: Among the 1873 detected features in IMS data, three lipid classes, including lysophosphatidylcholine (LysoPC), lysophosphatidylethanolamine (LysoPE) and lysophosphatidic acid (LysoPA) were observed nearly exclusively in SDD while presumed precursors, including phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidic acid (PA) lipids were detected in SDD and adjacent photoreceptor outer segments. Molecular signals specific to SDD were found in central retina and elsewhere. IHC results indicated abundant PLA2G5 in photoreceptors and retinal pigment epithelium (RPE). Discussion: The abundance of lysolipids in SDD implicates lipid remodeling or degradation in deposit formation, consistent with ultrastructural evidence of electron dense lipid-containing structures distinct from photoreceptor outer segment disks and immunolocalization of secretory PLA2G5 in photoreceptors and RPE. Further studies are required to understand the role of lipid signals observed in and around SDD.
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This paper reported the tensile failure strengths and damage procedure of composite laminate manufactured from the toughened-epoxy T800 prepreg at multi-scale levels. According to the exterior and interior distinction of each layer in laminate, the macro/mesoscale representative volume element (macro-RVE, meso-RVE) was first constructed, respectively. Then the micro-scale representative volume element (micro-RVE) with a hexagonal fiber-packed pattern in the interior zone of each layer in the laminate was finally determined on the principle of the same fiber volume fraction between the composite laminate and multi-scale RVEs. In the multi-RVEs analysis, the mechanical failure strengths of each scale model were transmitted from the last-scale model's homogenization, such as the meso-RVE from micro-RVE and the macro-RVE from meso-RVE. Based on our previous report, the innovative multi-scale damage and post-damage models on the concept of the smear crack were improved fully and incorporated by user-defined material subroutines (UMATs), such as in the addition of multiple cracks co-coupled, which makes it predict the element damage procedure. The averaged mechanical responses with damage mechanism of multi-scale RVEs under tensile, compressive, or shear loadings were obtained wholly by the homogenization method. The macroscale tensile damage initiation and propagation procedure were analyzed in detail including their global/local responses, being extended to comparison with experimental results.
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Thin-ply composite failure modes also significantly differ from conventional ply composite failure modes, with the final failure mechanism switching from irregular progressive failure to direct fracture characterized by a uniform fracture with the reduction of the ply thickness. When open holes and bolt joints are involved, thin-ply-laminated composites exhibit more complex stress states, damage evolution, and failure modes. Compared to the experimental study of thin-ply-laminated composite-bolted joints, there are few reports about numerical analysis. In order to understand the damage evolution and failure mechanism of thin-ply-laminated composites jointed by single-lap bolt, a progressive damage model based on three-dimensional (3D) LaRC failure criterion combined with cohesive element is constructed. Through an energy-based damage evolution method, this model can capture some significant mechanical characteristics in thin-ply-laminated structures, such as the in situ effect, delamination inhibition, and fiber compressive kinking failure. The comparisons between the numerical predictions and experimental observations are made to verify the accuracy of the proposed model. It is found that the predicted stress-displacement curves, failure modes, damage morphologies, etc., are consistent with the experimental results, indicating that the presented progressive damage analysis method displays excellent accuracy. The predicted stress at the onset of delamination is 50% higher than that of the conventional thick materials, which is also consistent with experimental results. Moreover, the numerical model provides evidence that the microstructure of thin-ply-laminated composite performs better in uniformity, which is more conducive to inhibiting the intra-layer damage and the expansion of delamination damage between layers. This study on the damage inhibition mechanism of thin-ply provides a potential analytical tool for evaluating damage tolerance and bearing capabilities in thin-ply-laminated composite-bolted joints.
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Purpose: The purpose of this study was to investigate histologic autofluorescence lifetimes and spectra of retinal pigment epithelium (RPE) on the transition from normal aging to RPE activation and migration in age-related macular degeneration (AMD). Methods: Autofluorescence lifetimes and spectra of 9 donor eyes were analyzed in cryosections by means of 2-photon excited fluorescence at 960 nm. Spectra were detected at 483 to 665 nm. Lifetimes were measured using time-correlated single photon counting in 2 spectral channels: 500 to 550 nm (short-wavelength spectral channel [SSC]) and 550 to 700 nm (long-wavelength spectral channel [LSC]). Fluorescence decays over time were approximated by a series of three exponential functions. The amplitude-weighted mean fluorescence lifetime was determined. Markers for retinoid activity (RPE65) and immune function (CD68) were immunolocalized in selected neighboring sections. Results: We identified 9 RPE morphology phenotypes resulting in 399 regions of interest (ROIs) for spectral and 497 ROIs for lifetime measurements. RPE dysmorphia results in a shorter wavelength peak of spectral emission: normal aging versus RPE migrated into the retina (intraELM) = 601.7 (9.5) nm versus 581.6 (7.3) nm, P < 0.001, whereas autofluorescence lifetimes increase: normal aging versus intraELM: SSC 180 (44) picosecond (ps) versus 320 (86) ps, P < 0.001; and LSC 250 (55) ps versus 441 (76) ps, P < 0.001. Ectopic RPE within the neurosensory retina is strongly CD68 positive and RPE65 negative. Conclusions: In the process of RPE degeneration, comprising different steps of dysmorphia and migration, lengthening of autofluorescence lifetimes and a hypsochromic shift of emission spectra can be observed. These autofluorescence changes might provide early biomarkers for AMD progression and contribute to our understanding of RPE-driven pathology.
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Degeneração Macular , Epitélio Pigmentado da Retina , Humanos , Epitélio Pigmentado da Retina/patologia , Oftalmoscopia/métodos , Degeneração Macular/diagnóstico , Degeneração Macular/patologia , Retina/patologia , Tomografia de Coerência Óptica/métodos , Angiofluoresceinografia/métodosRESUMO
To date there is no effective therapy for Alzheimer disease (AD). High levels of circulating high density lipoprotein (HDL) and its main protein, apolipoprotein A-I (apoA-I), reduce the risk of cardiovascular disease. Clinical studies show that plasma HDL cholesterol and apoA-I levels are low in patients with AD. To investigate if increasing plasma apoA-I/HDL levels ameliorates AD-like memory deficits and amyloid-ß (Aß) deposition, we generated a line of triple transgenic (Tg) mice overexpressing mutant forms of amyloid-ß precursor protein (APP) and presenilin 1 (PS1) as well as human apoA-I (AI). Here we show that APP/PS1/AI triple Tg mice have a 2-fold increase of plasma HDL cholesterol levels. When tested in the Morris water maze for spatial orientation abilities, whereas APP/PS1 mice develop age-related learning and memory deficits, APP/PS1/AI mice continue to perform normally during aging. Interestingly, no significant differences were found in the total level and deposition of Aß in the brains of APP/PS1 and APP/PS1/AI mice, but cerebral amyloid angiopathy was reduced in APP/PS1/AI mice. Also, consistent with the anti-inflammatory properties of apoA-I/HDL, glial activation was reduced in the brain of APP/PS1/AI mice. In addition, Aß-induced production of proinflammatory chemokines/cytokines was decreased in mouse organotypic hippocampal slice cultures expressing human apoA-I. Therefore, we conclude that overexpression of human apoA-I in the circulation prevents learning and memory deficits in APP/PS1 mice, partly by attenuating neuroinflammation and cerebral amyloid angiopathy. These findings suggest that elevating plasma apoA-I/HDL levels may be an effective approach to preserve cognitive function in patients with AD.
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Precursor de Proteína beta-Amiloide/fisiologia , Apolipoproteína A-I/metabolismo , Angiopatia Amiloide Cerebral/prevenção & controle , Transtornos Cognitivos/prevenção & controle , Modelos Animais de Doenças , Inflamação/prevenção & controle , Presenilina-1/fisiologia , Peptídeos beta-Amiloides , Animais , Apolipoproteína A-I/genética , Comportamento Animal , Western Blotting , Encéfalo/metabolismo , Encéfalo/patologia , Células Cultivadas , Angiopatia Amiloide Cerebral/etiologia , Colesterol/sangue , Transtornos Cognitivos/etiologia , Embrião de Mamíferos/citologia , Embrião de Mamíferos/metabolismo , Ensaio de Imunoadsorção Enzimática , Humanos , Técnicas Imunoenzimáticas , Inflamassomos , Inflamação/etiologia , Lipoproteínas/sangue , Aprendizagem em Labirinto , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Microglia , Mutação/genética , Miócitos de Músculo Liso/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Deleção de Sequência , Transdução de SinaisRESUMO
Compared with standard-ply composites, thin-ply composites exhibit a superior mechanical performance under various operating conditions due to their positive size effects. Thin-ply laminate failure modes, including matrix initial damage (MID), matrix failure (MF), and fiber failure (FF), have been distinguished through a systematic acoustic emission (AE) signals analysis combined with scanning electron microscopy (SEM). First, the characteristic frequencies of various failure modes are identified based on unidirectional laminates ([90] 68 and [0] 68). Then, according to the identified frequencies corresponding to distinctive damage modes, four lay-up sequences (02[[90m/0m]ns]02, m = 1, 2, 4, 8, n × m = 16) with a constant total thickness are designed, and the effects of the number of identical plies in the laminate thickness on the damage evolution characteristics and the damage process under uniaxial tension loads are dynamically monitored. The obtained results indicate that the characteristic frequency ranges for MID, MF, and FF are identified as 0-85 kHz, 165-260 kHz, and 261-304 kHz, respectively. The thickness of identical plies has a significant effect on onset damage. With the decrease of the number of identical plies (i.e., m in the stacking sequences), the thin-ply laminates exhibit the initiation of damage suppression effects and crack propagation resistance.
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Purpose: By optical coherence tomography (OCT) imaging, hyperreflective foci (HRF) indicate progression risk for advanced age-related macular degeneration (AMD) and are in part attributable to ectopic retinal pigment epithelium (RPE). We hypothesized that ectopic RPE are molecularly distinct from in-layer cells and that their cross-retinal course follows Müller glia. Methods: In clinical OCT (61 eyes, 44 patients with AMD, 79.4 ± 7.7 years; 29 female; follow-up = 4.7 ± 0.9 years), one HRF type, RPE plume (n = 129 in 4 morphologies), was reviewed. Twenty eyes of 20 donors characterized by ex vivo OCT were analyzed by histology (normal, 4; early/intermediate AMD, 7; geographic atrophy, 6; neovascular AMD, 3). Cryosections were stained with antibodies to retinoid (RPE65, CRALPB) and immune (CD68, CD163) markers. In published RPE cellular phenotypes, red immunoreactivity was assessed semiquantitatively by one observer (none, some cells, all cells). Results: Plume morphology evolved over time and many resolved (40%). Trajectories of RPE plume and cellular debris paralleled Müller glia, including near atrophy borders. RPE corresponding to HRF lost immunoreactivity for retinoid markers and gained immunoreactivity for immune markers. Aberrant immunoreactivity appeared in individual in-layer RPE cells and extended to all abnormal phenotypes. Müller glia remained CRALBP positive. Plume cells approached and contacted retinal capillaries. Conclusions: HRF are indicators not predictors of overall disease activity. Gain and loss of function starts with individual in-layer RPE cells and extends to all abnormal phenotypes. Evidence for RPE transdifferentiation, possibly due to ischemia, supports a proposed process of epithelial-mesenchyme transition. Data can propel new biomarkers and therapeutic strategies for AMD.
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Angiofluoresceinografia/métodos , Refração Ocular/fisiologia , Epitélio Pigmentado da Retina/patologia , Tomografia de Coerência Óptica/métodos , Degeneração Macular Exsudativa/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Transdiferenciação Celular , Progressão da Doença , Feminino , Seguimentos , Fundo de Olho , Humanos , Masculino , Prognóstico , Fatores de Tempo , Degeneração Macular Exsudativa/fisiopatologiaRESUMO
Recent evidence indicates that inflammation may significantly contribute to the pathogenesis of Alzheimer's disease (AD). Since the apo A-I mimetic peptide D-4F has been shown to inhibit atherosclerotic lesion formation and regress already existing lesions (in the presence of pravastatin) and the peptide also decreases brain arteriole inflammation, we undertook a study to evaluate the efficacy of oral D-4F co-administered with pravastatin on cognitive function and amyloid beta (A beta) burden in the hippocampus of APPSwe-PS1 Delta E9 mice. Three groups of male mice were administered D-4F and pravastatin, Scrambled D-4F (ScD-4F, a control peptide) and pravastatin in drinking water, while drinking water alone served as control. The escape latency in the Morris Water Maze test was significantly shorter for the D-4F+statin administered animals compared to the other two groups. While the hippocampal region of the brain was covered with 4.2+/-0.5 and 3.8+/-0.6% of A beta load in the control and ScD-4F+statin administered groups, in the D-4F+statin administered group A beta load was only 1.6+/-0.1%. Furthermore, there was a significant decrease in the number of activated microglia (p<0.05 vs the other two groups) and activated astrocytes (p<0.05 vs control) upon oral D-4F+statin treatment. Inflammatory markers TNFalpha and IL-1 beta levels were decreased significantly in the D-4F+statin group compared to the other two groups (for IL-1 beta p<0.01 vs the other two groups and for TNF-alpha p<0.001 vs control) and the expression of MCP-1 were also less in D-4F+statin administered group compared to the other two groups. These results suggest that the apo A-I mimetic peptide inhibits amyloid beta deposition and improves cognitive function via exerting anti-inflammatory properties in the brain.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides/metabolismo , Apolipoproteína A-I/administração & dosagem , Cognição/efeitos dos fármacos , Hipocampo/metabolismo , Administração Oral , Doença de Alzheimer/complicações , Doença de Alzheimer/metabolismo , Doença de Alzheimer/psicologia , Precursor de Proteína beta-Amiloide/genética , Animais , Anticolesterolemiantes/administração & dosagem , Astrócitos/efeitos dos fármacos , Astrócitos/fisiologia , Quimiocina CCL2/metabolismo , Colesterol/sangue , Modelos Animais de Doenças , Quimioterapia Combinada , Hipocampo/efeitos dos fármacos , Hipocampo/fisiopatologia , Interleucina-1beta/metabolismo , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Camundongos , Camundongos Transgênicos , Microglia/efeitos dos fármacos , Microglia/fisiologia , Placa Amiloide/efeitos dos fármacos , Placa Amiloide/fisiologia , Pravastatina/administração & dosagem , Nexinas de Proteases , Receptores de Superfície Celular/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Conflicting results have been reported on the association of plasma total homocysteine (tHcy) and cholesterol levels in Alzheimer disease (AD). The objective of this study was to determine the relationship between cognitive performance and plasma levels of tHcy and its biological determinants folate and vitamin B(12), and lipids in clinically diagnosed AD patients. METHODS: A cross-sectional database review was performed on two separate groups of patients (n = 191). Mini-Mental State Exam (MMSE) scores, plasma levels of tHcy, vitamin B(12), folate, cholesterol, and triglycerides were analyzed. RESULTS: The MMSE scores were inversely correlated with age, plasma levels of tHcy and LDL cholesterol. However, only the inverse relationship between MMSE scores and LDL cholesterol levels persisted after adjustment for age, sex, and status of statin treatment. Plasma tHcy levels increased significantly with age and were inversely related to vitamin B(12) and folate levels, which modified the relationship between MMSE scores and plasma tHcy levels. CONCLUSIONS: The plasma tHcy levels appeared to relate more to aging than to cognition. Cognitive performance was inversely associated with plasma LDL cholesterol levels in AD patients. Our findings provide further evidence that high LDL cholesterol levels may play a role in the pathogenesis of AD.
Assuntos
Doença de Alzheimer/sangue , Doença de Alzheimer/psicologia , Cognição/fisiologia , Ácido Fólico/sangue , Homocisteína/sangue , Lipídeos/sangue , Desempenho Psicomotor/fisiologia , Vitamina B 12/sangue , Idoso , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Estudos Transversais , Bases de Dados Factuais , Feminino , Humanos , Masculino , Caracteres Sexuais , Triglicerídeos/sangueRESUMO
We recently proposed an all-atom model for apolipoprotein (apo) A-I in discoidal high-density lipoprotein in which two monomers form stacked antiparallel helical rings rotationally aligned by interhelical salt-bridges. The model can be derived a priori from the geometry of a planar bilayer disc that constrains the hydrophobic face of a continuous amphipathic alpha helix in lipid-associated apoA-I to a plane inside of an alpha-helical torus. This constrains each apoA-I monomer to a novel conformation, that of a slightly unwound, curved, planar amphipathic alpha 11/3 helix (three turns per 11 residues). Using non-denaturing gradient gel electrophoresis, we show that dimyristoylphosphocholine discs containing two apoA-I form five distinct particles with maximal Stokes diameters of 98 A (R2-1), 106 A (R2-2), 110 A (R2-3), 114 A (R2-4) and 120 A (R2-5). Further, we show that the Stokes diameters of R2-1 and R2-2 are independent of the N-terminal 43 residues (the flexible domain) of apoA-I, while the flexible domain is necessary and sufficient for the formation of the three larger complexes. On the basis of these results, the conformation of apoA-I on the R2-2 disc can be modeled accurately as an amphipathic helical double belt extending the full length of the lipid-associating domain with N and C-terminal ends in direct contact. The smallest of the discs, R2-1, models as the R2-2 conformation with an antiparallel 15-18 residue pairwise segment of helixes hinged off the disc edge. The conformations of full-length apoA-I on the flexible domain-dependent discs (R2-3, R2-4 and R2-5) model as the R2-2 conformation extended on the disc edge by one, two or three of the 11-residue tandem amphipathic helical repeats (termed G1, G2 and G3), respectively, contained within the flexible domain. Although we consider these results to favor the double belt model, the topographically very similar hairpin-belt model cannot be ruled out entirely.