RESUMEN
BACKGROUND: Varicose veins affect approximately 25% of people in industrialized countries. METHODS: The study aimed at detecting apoptotic cells and histopathological changes in varicose vein walls. Patients (N.=41) with varicose veins and 30 control group patients were divided into two groups according to their age (younger and older than 50 years). Apoptosis was determined by the TUNEL assay, elastin and collagen IV expression by immunohistochemistry and ultrastructural changes by transmission electron microscopy. RESULTS: The results show that the number of apoptotic cells in the layers of varicose veins increased, in particular in a group of patients aged over 50 years. In the varicose veins as compared to control veins the elastic fibers were found to be thinner, more fragmented and disorderly arranged. Elastin and collagen IV expression was found to decline in the intima and the media of varicose veins in both age groups. Electron microscopy demonstrated hypertrophy and degeneration of smooth muscle cells. Furthermore, cells with ultrastructural feature of apoptosis were noted. In the disorganized and expanded extracellular matrix membrane-bound vesicles, ghost bodies with different size and electron density were observed. Ghost bodies seem to bud off from smooth muscle cells and are likely to be involved in extracellular matrix remodeling as they are seen in close contact with collagen fibers. CONCLUSIONS: The study demonstrates increase of apoptotic cells in the wall of varicose veins along with vein wall structural abnormalities including alterations of smooth muscle cells and decline of elastin and collagen IV expression.
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Apoptosis , Elastina , Microscopía Electrónica de Transmisión , Miocitos del Músculo Liso , Vena Safena , Várices , Humanos , Vena Safena/ultraestructura , Vena Safena/patología , Vena Safena/metabolismo , Persona de Mediana Edad , Elastina/metabolismo , Várices/patología , Várices/metabolismo , Femenino , Adulto , Masculino , Miocitos del Músculo Liso/ultraestructura , Miocitos del Músculo Liso/patología , Miocitos del Músculo Liso/metabolismo , Anciano , Estudios de Casos y Controles , Colágeno Tipo IV/metabolismo , Músculo Liso Vascular/ultraestructura , Músculo Liso Vascular/patología , Músculo Liso Vascular/metabolismo , Inmunohistoquímica , Insuficiencia Venosa/patología , Insuficiencia Venosa/metabolismo , Adulto Joven , Factores de Edad , Tejido Elástico/ultraestructura , Tejido Elástico/metabolismo , Tejido Elástico/patologíaRESUMEN
Two major constituents of exfoliation material, fibrillin-1 and lysyl oxidase-like 1 (encoded by FBN1 and LOXL1), are implicated in exfoliation glaucoma, yet their individual contributions to ocular phenotype are minor. To test the hypothesis that a combination of FBN1 mutation and LOXL1 deficiency exacerbates ocular phenotypes, the pan-lysyl oxidase inhibitor ß-aminopropionitrile (BAPN) was used to treat adult wild-type (WT) mice and mice heterozygous for a missense mutation in Fbn1 (Fbn1C1041G/+) for 8 weeks and their eyes were examined. Although intraocular pressure did not change and exfoliation material was not detected in the eyes, BAPN treatment worsened optic nerve and axon expansion in Fbn1C1041G/+ mice, an early sign of axonal damage in rodent models of glaucoma. Disruption of elastic fibers was detected only in Fbn1C1041G/+ mice, which increased with BAPN treatment, as shown by histologic and immunohistochemical staining of the optic nerve pia mater. Transmission electron microscopy showed that Fbn1C1041G/+ mice had fewer microfibrils, smaller elastin cores, and a lower density of elastic fibers compared with WT mice in control groups. BAPN treatment led to elastin core expansion in both WT and Fbn1C1041G/+ mice, but an increase in the density of elastic fiber was confined to Fbn1C1041G/+ mice. LOX inhibition had a stronger effect on optic nerve and elastic fiber parameters in the context of Fbn1 mutation, indicating the Marfan mouse model with LOX inhibition warrants further investigation for exfoliation glaucoma pathogenesis.
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Aminopropionitrilo , Modelos Animales de Enfermedad , Fibrilina-1 , Síndrome de Marfan , Nervio Óptico , Proteína-Lisina 6-Oxidasa , Animales , Ratones , Adipoquinas , Aminoácido Oxidorreductasas/metabolismo , Aminoácido Oxidorreductasas/antagonistas & inhibidores , Aminoácido Oxidorreductasas/genética , Aminopropionitrilo/farmacología , Tejido Elástico/patología , Tejido Elástico/metabolismo , Tejido Elástico/ultraestructura , Fibrilina-1/genética , Fibrilinas/metabolismo , Glaucoma/patología , Presión Intraocular , Síndrome de Marfan/patología , Síndrome de Marfan/complicaciones , Ratones Endogámicos C57BL , Proteínas de Microfilamentos/metabolismo , Nervio Óptico/patología , Nervio Óptico/ultraestructura , Nervio Óptico/efectos de los fármacos , Proteína-Lisina 6-Oxidasa/metabolismo , Proteína-Lisina 6-Oxidasa/antagonistas & inhibidoresRESUMEN
OBJECTIVE: Aortic aneurysm and dissection are major life-threatening complications of Marfan syndrome. Avoiding factors that promote aortic damage is critical in managing the care of these patients. Findings from clinical and animal studies raise concerns regarding fluoroquinolone use in patients at risk for aortic aneurysm and dissection. Therefore, we examined the effects of ciprofloxacin on aortic aneurysm and dissection development in Marfan mice. METHODS: Eight-week-old Marfan mice (Fbn1C1041G/+) were given ciprofloxacin (100 mg/kg/d; n = 51) or vehicle (n = 59) for 4 weeks. Mice were monitored for 16 weeks. Aortic diameters were measured by using ultrasonography, and aortic structure was examined by using histopathologic and immunostaining analyses. RESULTS: Vehicle-treated Fbn1C1041G/+ mice showed progressive aortic enlargement, with aortic rupture occurring in 5% of these mice. Compared with vehicle-treated Fbn1C1041G/+ mice, ciprofloxacin-treated Fbn1C1041G/+ mice showed accelerated aortic enlargement (P = .01) and increased incidences of aortic dissection (25% vs 47%, P = .03) and rupture (5% vs 25%, P = .005). Furthermore, ciprofloxacin-treated Fbn1C1041G/+ mice had higher levels of elastic fiber fragmentation, matrix metalloproteinase expression, and apoptosis than did vehicle-treated Fbn1C1041G/+ mice. CONCLUSIONS: Ciprofloxacin accelerates aortic root enlargement and increases the incidence of aortic dissection and rupture in Marfan mice, partially by suppressing lysyl oxidase expression and further compromising the inherited defect in aortic elastic fibers. Our findings substantiate that ciprofloxacin should be avoided in patients with Marfan syndrome.
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Antibacterianos/toxicidad , Aorta/efectos de los fármacos , Aneurisma de la Aorta/inducido químicamente , Disección Aórtica/inducido químicamente , Rotura de la Aorta/inducido químicamente , Ciprofloxacina/toxicidad , Fibrilina-1/genética , Remodelación Vascular/efectos de los fármacos , Disección Aórtica/genética , Disección Aórtica/metabolismo , Disección Aórtica/patología , Animales , Aorta/metabolismo , Aorta/ultraestructura , Aneurisma de la Aorta/genética , Aneurisma de la Aorta/metabolismo , Aneurisma de la Aorta/patología , Rotura de la Aorta/genética , Rotura de la Aorta/metabolismo , Rotura de la Aorta/patología , Apoptosis/efectos de los fármacos , Dilatación Patológica , Progresión de la Enfermedad , Tejido Elástico/efectos de los fármacos , Tejido Elástico/metabolismo , Tejido Elástico/ultraestructura , Proteínas de la Matriz Extracelular/metabolismo , Femenino , Predisposición Genética a la Enfermedad , Masculino , Metaloproteinasas de la Matriz/metabolismo , Ratones Noqueados , Fenotipo , Proteína-Lisina 6-Oxidasa/metabolismoRESUMEN
OBJECTIVE: Aneurysmal subarachnoid hemorrhage remains a devastating event with poorly understood pathophysiology. Previous studies have suggested that aneurysm wall inflammation may play a part in the development and potential rupture of aneurysms. The rabbit elastase aneurysm model is a well-established model, which produces aneurysms closely mimicking human cerebral aneurysms in flow dynamics and histopathology. The primary aim of this study was to correlate inflammatory changes after aneurysm formation using sequential vessel wall imaging with histopathologic analysis. A secondary aim was to evaluate the potential effect of gender and anti-inflammatory treatment with aspirin on this inflammatory response. METHODS: Twenty-seven New Zealand rabbits underwent surgery to create an aneurysm using elastase infusion at the right common carotid artery origin. Vessel wall imaging and histopathologic analysis was obtained at different time points after aneurysm creation. The rabbits were also randomized by gender and to treatment groups with or without aspirin. RESULTS: Histopathologic analysis revealed 3 distinct phases after aneurysm formation. These phases were an initial inflammatory phase, followed by a regeneration phase, and finally a connective tissue deposition phase. Vessel wall imaging demonstrated 2 distinct imaging patterns. No appreciable differences were seen in histology or imaging when comparing gender or treatment with aspirin. CONCLUSIONS: Inflammatory changes induced by the rabbit elastase aneurysm model can be correlated with histopathologic findings and observed on noninvasive vessel wall imaging. This may provide a method to study the inflammatory pathway as it pertains to aneurysmal development and subsequent rupture.
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Enfermedades de las Arterias Carótidas/inducido químicamente , Modelos Animales de Enfermedad , Aneurisma Intracraneal/complicaciones , Angiografía por Resonancia Magnética , Elastasa Pancreática/toxicidad , Conejos/fisiología , Animales , Aspirina/uso terapéutico , Enfermedades de las Arterias Carótidas/diagnóstico por imagen , Enfermedades de las Arterias Carótidas/patología , Arteria Carótida Común/diagnóstico por imagen , Arteria Carótida Común/efectos de los fármacos , Arteria Carótida Común/patología , Arteria Carótida Común/fisiología , Progresión de la Enfermedad , Tejido Elástico/ultraestructura , Femenino , Hiperplasia , Infusiones Intraarteriales , Aneurisma Intracraneal/inducido químicamente , Aneurisma Intracraneal/diagnóstico por imagen , Aneurisma Intracraneal/tratamiento farmacológico , Masculino , Miocitos del Músculo Liso/patología , Necrosis , Elastasa Pancreática/administración & dosificación , Conejos/inmunología , Regeneración , Caracteres Sexuales , Método Simple Ciego , Túnica Íntima/patología , Túnica Media/patología , Vasculitis/tratamiento farmacológico , Vasculitis/etiología , Vasculitis/patologíaRESUMEN
Supramolecular networks composed of fibrillins (fibrillin-1 and fibrillin-2) and associated ligands form intricate cellular microenvironments which balance skin homoeostasis and direct remodelling. Fibrillins assemble into microfibrils which are not only indispensable for conferring elasticity to the skin, but also control the bioavailability of growth factors targeted to the extracellular matrix architecture. Fibrillin microfibrils (FMF) represent the core scaffolds for elastic fibre formation, and they also decorate the surface of elastic fibres and form independent networks. In normal dermis, elastic fibres are suspended in a three-dimensional basket-like lattice of FMF intersecting basement membranes at the dermal-epidermal junction and thus conferring pliability to the skin. The importance of FMF for skin homoeostasis is illustrated by the clinical features caused by mutations in the human fibrillin genes (FBN1, FBN2), summarized as "fibrillinopathies." In skin, fibrillin mutations result in phenotypes ranging from thick, stiff and fibrotic skin to thin, lax and hyperextensible skin. The most plausible explanation for this spectrum of phenotypic outcomes is that FMF regulate growth factor signalling essential for proper growth and homoeostasis of the skin. Here, we will give an overview about the current understanding of the underlying pathomechanisms leading to fibrillin-dependent fibrosis as well as forms of cutis laxa caused by mutational inactivation of FMF-associated ligands.
Asunto(s)
Enfermedades del Tejido Conjuntivo/genética , Tejido Elástico/metabolismo , Fibrilinas/genética , Fibrilinas/metabolismo , Homeostasis , Piel/metabolismo , Animales , Enfermedades del Desarrollo Óseo/genética , Tejido Elástico/ultraestructura , Elasticidad , Fibrilinas/ultraestructura , Fibrosis , Humanos , Deformidades Congénitas de las Extremidades/genética , Microfibrillas/metabolismo , Microfibrillas/ultraestructura , Conformación Molecular , Transducción de Señal , Piel/patología , Piel/ultraestructura , Fenómenos Fisiológicos de la Piel , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Purpose: To investigate the initial events in the development of the human cornea, focusing on cell migration, and extracellular matrix synthesis and organization. To determine whether elastic fibers are present in the extracellular matrix during early human corneal development. Methods: Human corneas were collected from week 7 to week 17 of development. An elastic fiber-enhancing stain, tannic acid-uranyl acetate, was applied to all tissue. Three-dimensional serial block-face scanning electron microscopy combined with conventional transmission electron microscopy was used to analyze the corneal stroma. Results: An acellular collagenous primary stroma with an orthogonal arrangement of fibrils was identified in the central cornea from week 7 of corneal development. At week 7.5, mesenchymal cells migrated toward the central cornea and associated with the acellular collagenous matrix. Novel cell extensions from the endothelium were identified. Elastic fibers were found concentrated in the posterior peripheral corneal stroma from week 12 of corneal development. Conclusions: This study provides novel evidence of an acellular primary stroma in the early development of the embryonic human cornea. Cell extensions exist as part of a communication system and are hypothesized to assist in the migration of the mesenchymal cells and the development of the mature cornea. Elastic fibers identified in early corneal development may play an important role in establishing corneal shape.
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Córnea/embriología , Sustancia Propia/embriología , Tejido Elástico/embriología , Endotelio Corneal/embriología , Movimiento Celular/fisiología , Córnea/ultraestructura , Sustancia Propia/ultraestructura , Tejido Elástico/ultraestructura , Endotelio Corneal/ultraestructura , Matriz Extracelular/ultraestructura , Edad Gestacional , Humanos , Imagenología Tridimensional , Microscopía Electrónica de Rastreo , Microscopía Electrónica de TransmisiónRESUMEN
Elastic fibers are an essential component of the extracellular matrix of connective tissues. The focus of both clinical management and scientific investigation of elastic fiber disorders has centered on the cardiovascular manifestations due to their significant impact on morbidity and mortality. As such, the current understanding of the orthopedic conditions experienced by these patients is limited. The musculoskeletal implications of more subtle elastic fiber abnormalities, whether due to allelic variants or age-related tissue degeneration, are also not well understood. Recent advances have begun to uncover the effects of elastic fiber deficiency on tendon and ligament biomechanics; future research must further elucidate mechanisms governing the role of elastic fibers in these tissues. The identification of population-based genetic variations in elastic fibers will also be essential. Minoxidil administration, modulation of protein expression with micro-RNA molecules, and direct injection of recombinant elastic fiber precursors have demonstrated promise for therapeutic intervention, but further work is required prior to consideration for orthopedic clinical application. This review provides an overview of the role of elastic fibers in musculoskeletal tissue, summarizes current knowledge of the orthopedic manifestations of elastic fiber abnormalities, and identifies opportunities for future investigation and clinical application.
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Tejido Elástico/fisiología , Fenómenos Fisiológicos Musculoesqueléticos , Animales , Fenómenos Biomecánicos , Enfermedades del Tejido Conjuntivo/genética , Tejido Elástico/ultraestructura , Humanos , Mecanotransducción Celular , OrtopediaRESUMEN
BACKGROUND: Aplasia cutis congenita (ACC) is a rare and heterogeneous disorder characterized by congenital absence of skin. The scalp is the most commonly affected site and lesions may overlie deeper ectodermal abnormalities. The exact etiology is still unknown, and histopathologic features are poorly defined. METHODS: A series of 10 cases from nine patients was analyzed to characterize the clinicopathologic spectrum and age-related changes of ACC of the scalp. Hematoxylin and eosin, S100, Elastica van Gieson, and Weigert elastic stains were performed, and clinical information was retrieved from archived medical files. RESULTS: Patient ages ranged from 1 day to 39 years (median 57 months). All cases resembled deep-reaching scars with almost complete loss of all adnexal structures. Isolated residual hair follicles were present in 8/10 and sweat glands and ducts in 2/10 cases. The subcutis was thinned or absent. Elastic fibers were always more fragmented than in normal tissue, and the thickness and density increased over time. There was no gain of adnexal structures with increasing age. CONCLUSIONS: ACC represents a congenital scarring alopecia with permanent loss of skin appendages. Histopathologic changes resemble a deep-reaching scar with fragmented elastic fibers and differentiate ACC from all other forms of non-traumatic congenital alopecias.
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Displasia Ectodérmica/patología , Tejido Elástico/patología , Neoplasias de Anexos y Apéndices de Piel/patología , Cuero Cabelludo/patología , Adolescente , Adulto , Niño , Preescolar , Cicatriz/patología , Tejido Elástico/ultraestructura , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Neoplasias de Anexos y Apéndices de Piel/diagnóstico , Estudios Retrospectivos , Proteínas S100/metabolismo , Cuero Cabelludo/anomalías , Adulto JovenRESUMEN
Variable feather overlap enables birds to morph their wings, unlike aircraft. They accomplish this feat by means of elastic compliance of connective tissue, which passively redistributes the overlapping flight feathers when the skeleton moves to morph the wing planform. Distinctive microstructures form "directional Velcro," such that when adjacent feathers slide apart during extension, thousands of lobate cilia on the underlapping feathers lock probabilistically with hooked rami of overlapping feathers to prevent gaps. These structures unlock automatically during flexion. Using a feathered biohybrid aerial robot, we demonstrate how both passive mechanisms make morphing wings robust to turbulence. We found that the hooked microstructures fasten feathers across bird species except silent fliers, whose feathers also lack the associated Velcro-like noise. These findings could inspire innovative directional fasteners and morphing aircraft.
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Columbidae/anatomía & histología , Columbidae/fisiología , Plumas/ultraestructura , Vuelo Animal , Alas de Animales/ultraestructura , Animales , Tejido Elástico/fisiología , Tejido Elástico/ultraestructuraRESUMEN
According to our previous reports, the intraperiodontal elastic fiber system comprises oxytalan fibers, whereas all types of elastic system fibers are present in the gingiva. Much remains to be elucidated regarding the topographic development of the elastic fiber system that constitutes the walls of the digestive organs. This study aimed to examine the topographic development of the elastic fiber system in the periodontal tissue, oral cavity and digestive tract of rats at light- and electron microscopic levels. At embryonic day 20, in situ hybridization revealed the mRNA expression of tropoelastin in the putative gingival lamina propria but not in the dental follicle. At the postnatal stage, the masticatory mucous membrane of the gingiva and hard palate comprised three different types of elastic system fibers (oxytalan, elaunin and elastic fibers). Conversely, the elastic fiber system comprised elaunin and elastic fibers in other oral mucosae and the lining mucosae of digestive tract organs (the esophagus, stomach and small intestine). The findings of our study suggest that the elastic fiber system is mainly related to tissue resistance in the periodontal ligament and tissue elasticity in the oral mucosae without masticatory mucosae and the overlying mucosa of digestive tracts and both functions in the gingiva and hard palate, respectively. The appearance of elaunin fibers in the periodontium of rats aged 14 weeks suggests the expression of tropoelastin induced by mechanical stressors such as mastication. The intraperiodontal difference in the distribution of elaunin fibers suggests heterogeneity among fibroblasts constituting the periodontium.
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Tejido Elástico/ultraestructura , Tracto Gastrointestinal/citología , Tracto Gastrointestinal/ultraestructura , Boca/ultraestructura , Animales , Proteínas Contráctiles/genética , Tejido Elástico/química , Femenino , Masculino , Microscopía , Microscopía Electrónica , Boca/citología , Ligamento Periodontal/química , Ligamento Periodontal/citología , Ligamento Periodontal/ultraestructura , Ratas , Ratas Wistar , Coloración y Etiquetado , Tropoelastina/genéticaRESUMEN
RATIONALE: Elastin is an important ECM (extracellular matrix) protein in large and small arteries. Vascular smooth muscle cells (SMCs) produce the layered elastic laminae found in elastic arteries but synthesize little elastin in muscular arteries. However, muscular arteries have a well-defined internal elastic lamina (IEL) that separates endothelial cells (ECs) from SMCs. The extent to which ECs contribute elastin to the IEL is unknown. OBJECTIVE: To use targeted elastin (Eln) deletion in mice to explore the relative contributions of SMCs and ECs to elastic laminae formation in different arteries. METHODS AND RESULTS: We used SMC- and EC-specific Cre recombinase transgenes with a novel floxed Eln allele to focus gene inactivation in mice. Inactivation of Eln in SMCs using Sm22aCre resulted in depletion of elastic laminae in the arterial wall with the exception of the IEL and SMC clusters in the outer media near the adventitia. Inactivation of elastin in ECs using Tie2Cre or Cdh5Cre resulted in normal medial elastin and a typical IEL in elastic arteries. In contrast, the IEL was absent or severely disrupted in muscular arteries. Interruptions in the IEL resulted in neointimal formation in the ascending aorta but not in muscular arteries. CONCLUSIONS: Combined with lineage-specific fate mapping systems, our knockout results document an unexpected heterogeneity in vascular cells that produce the elastic laminae. SMCs and ECs can independently form an IEL in most elastic arteries, whereas ECs are the major source of elastin for the IEL in muscular and resistance arteries. Neointimal formation at IEL disruptions in the ascending aorta confirms that the IEL is a critical physical barrier between SMCs and ECs in the large elastic arteries. Our studies provide new information about how SMCs and ECs contribute elastin to the arterial wall and how local elastic laminae defects may contribute to cardiovascular disease.
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Tejido Elástico/metabolismo , Elastina/metabolismo , Células Endoteliales/metabolismo , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , Animales , Arterias/crecimiento & desarrollo , Arterias/metabolismo , Presión Sanguínea , Linaje de la Célula , Proliferación Celular , Tejido Elástico/crecimiento & desarrollo , Tejido Elástico/ultraestructura , Elastina/deficiencia , Elastina/genética , Células Endoteliales/ultraestructura , Femenino , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Liso Vascular/crecimiento & desarrollo , Músculo Liso Vascular/ultraestructura , Miocitos del Músculo Liso/ultraestructura , Neointima , Transducción de SeñalRESUMEN
PURPOSE: To evaluate changes in the quantity of elastic fibers in the corpora cavernosa of rats during the natural aging process, and to assess the degree of this change by determining volumetric density (Vv) at different ages via stereological analysis. METHODS: Forty-eight rats, raised under similar conditions, were subjected to the natural aging process and divided into four groups (G1 to G4), according to age at the time of penectomy (6, 9, 12, and 24 months, respectively). Histological sections of the middle segment of the penis were stained with Weigert's resorcin-fuchsin, and the volumetric density (Vv) of elastic fibers of the corpora cavernosa were determined via stereological analysis. RESULTS: There were no statistically significant differences in Vv among groups G1, G2, and G3. These three groups were therefore considered as a single group. The mean Vv of this group showed a statistically significant reduction compared to that of G4 (0.16 vs. 0.11, p<0.05). CONCLUSION: Natural aging in rats was responsible for a reduction in volumetric density of elastic fibers of the corpora cavernosa (approximately 30% decrease in Vv) during senescence.
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Envejecimiento/fisiología , Tejido Elástico/ultraestructura , Células Endoteliales/fisiología , Pene/citología , Envejecimiento/patología , Animales , Colágeno/fisiología , Colágeno/ultraestructura , Tejido Elástico/patología , Tejido Elástico/fisiología , Disfunción Eréctil/fisiopatología , Masculino , Modelos Animales , Pene/fisiología , Ratas , Ratas WistarRESUMEN
The present study aims to identify specific staining methods for acoustic histology. We compared attenuation-of-sound (AOS) images from scanning acoustic microscopy (SAM) with light microscopy (LM) images. Ethanol-fixed tissue or cytology samples and formalin-fixed surgical or autopsy specimens were examined. Nuclei, collagen, elastic fibers and polysaccharides and various antigens, including cell surface, cytoplasmic, nuclear and stromal substances, were observed. Samples with various fixation methods were used. Hematoxylin staining had significantly higher AOS values in accordance with staining duration. Specific staining for collagen, elastic fibers and polysaccharides increased the AOS values of the specific substance. Using diaminobenzidine tetrahydrochloride in NiCl2 solution as a substrate for horseradish peroxidase increased the AOS values to those suitable for acoustic immunostaining. Collagenase digestion after collagen staining decreased AOS values, reflecting collagen density and distribution. Staining with specific dyes or acoustic immunostaining enabled the histologic localization of specific substances by SAM, similar to LM.
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Anticuerpos/ultraestructura , Colágeno/ultraestructura , Colorantes , Tejido Elástico/ultraestructura , Microscopía Acústica/métodos , Polisacáridos/ultraestructura , Cadáver , Humanos , Inmunohistoquímica , Estudios RetrospectivosRESUMEN
Elastin is an essential structural protein in the extracellular matrix of vertebrates. It is the core component of elastic fibers, which enable connective tissues such as those of the skin, lungs or blood vessels to stretch and recoil. This function is provided by elastin's exceptional properties, which mainly derive from a unique covalent cross-linking between hydrophilic lysine-rich motifs of units of the monomeric precursor tropoelastin. To date, elastin's cross-linking is poorly investigated. Here, we purified elastin from human tissue and cleaved it into soluble peptides using proteases with different specificities. We then analyzed elastin's molecular structure by identifying unmodified residues, post-translational modifications and cross-linked peptides by high-resolution mass spectrometry and amino acid analysis. The data revealed the presence of multiple isoforms in parallel and a complex and heterogeneous molecular interconnection. We discovered that the same lysine residues in different monomers were simultaneously involved in various cross-link types or remained unmodified. Furthermore, both types of cross-linking domains, Lys-Pro and Lys-Ala domains, participate not only in bifunctional inter- but also in intra-domain cross-links. We elucidated the sequences of several desmosine-containing peptides and the contribution of distinct domains such as 6, 14 and 25. In contrast to earlier assumptions proposing that desmosine cross-links are formed solely between two domains, we elucidated the structure of a peptide that proves a desmosine formation with participation of three Lys-Ala domains. In summary, these results provide new and detailed insights into the cross-linking process, which takes place within and between human tropoelastin units in a stochastic manner.
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Elastina/química , Lisina/química , Péptidos/química , Tropoelastina/química , Secuencia de Aminoácidos/genética , Desmosina/química , Tejido Elástico/química , Tejido Elástico/ultraestructura , Elastina/ultraestructura , Matriz Extracelular/química , Matriz Extracelular/ultraestructura , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Espectrometría de Masas , Estructura Molecular , Isoformas de Proteínas/química , Isoformas de Proteínas/ultraestructura , Procesamiento Proteico-Postraduccional/genética , Piel/química , Tropoelastina/ultraestructuraRESUMEN
PURPOSE: The objective of this study was to analyze the layers of yellow ligament in lumbar canal stenosis and disk herniation. METHODS: Eighteen ligaments were harvested from patients with lumbar spinal canal stenosis. Twenty-nine normal samples from lumbar spine disk herniation patients served as control. All surgical procedures were the same. Ligaments were stained in hematoxylin and eosin; picrosirius-hematoxylin for collagen; Weigert's resorcin-fuchsin for elaunin, oxytalan and elastic fibers; and transmission electron microscopy. Immunohistochemistry was performed for Il-6; Il-10; and CD-31, PGP9.5. Results are described in means and standard error (mean ± SE), and all analyses adopted the significance level of P < 0.05. RESULTS: Spinal stenosis ligaments were 2.5 × thicker. Control superficial ligaments presented a large number of thick, compact collagen fibers and a significant amount of oxytalan and mature elastic fibers. The deep layer presented a large number of mature elastic fibers. In the stenosis group, collagen was thinner and compacted in both layers. There was no difference in the interleukin profile among groups. The deep portion of the stenosis group presented a higher number of vessels and nerves. CONCLUSION: Two layers compose the elastic system of the normal ligamentum flavum, where the deep portion is mainly responsible for its elasticity (elaunin fibers), while its resistance depends on the concentration of oxytalan fibers, which are more present in the superficial layer. Ligamentum flavum in the stenosis samples presents more mononuclear infiltrate and more degraded elastic fibers with a higher number of vessels in its deep portion. These slides can be retrieved under Electronic Supplementary Material.
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Degeneración del Disco Intervertebral/metabolismo , Ligamento Amarillo/química , Vértebras Lumbares/química , Estenosis Espinal/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Proteínas Contráctiles/análisis , Tejido Elástico/química , Tejido Elástico/patología , Tejido Elástico/ultraestructura , Elasticidad , Proteínas de la Matriz Extracelular/análisis , Femenino , Humanos , Degeneración del Disco Intervertebral/patología , Desplazamiento del Disco Intervertebral/metabolismo , Desplazamiento del Disco Intervertebral/patología , Ligamento Amarillo/ultraestructura , Vértebras Lumbares/patología , Masculino , Microscopía Electrónica , Persona de Mediana Edad , Estenosis Espinal/patología , Adulto JovenRESUMEN
Individuals affected with autosomal recessive cutis laxa type 2B and 3 usually show translucent skin with visible veins and abnormal elastic fibers, intrauterine and/or postnatal growth restriction and a typical triangular facial gestalt. Here we describe three unrelated individuals in whom such a cutis laxa syndrome was suspected, especially after electron microscopy revealed immature and less dense dermal elastic fibers in one of them. However, one of these children also displayed optic atrophy and two hypogammaglobulinemia. All had elevated liver enzymes and acute liver failure during febrile episodes leading to early demise in two of them. The only surviving patient had been treated with immunoglobulins. Through exome sequencing we identified mutations in NBAS, coding for a protein involved in Golgi-to-ER transport. NBAS deficiency causes several rare conditions ranging from isolated recurrent acute liver failure to a multisystem disorder mainly characterized by short stature, optic nerve atrophy and Pelger-Huët anomaly (SOPH). Since we subsequently verified Pelger-Huët anomaly in two of the patients the diagnosis SOPH syndrome was unequivocally proven. Our data show that SOPH syndrome can be regarded as a differential diagnosis for the progeroid forms of cutis laxa in early infancy and that possibly treatment of the hypogammaglobulinemia can be of high relevance for the prognosis.
Asunto(s)
Trastornos del Crecimiento/diagnóstico , Proteínas de Neoplasias/genética , Enfermedades del Nervio Óptico/diagnóstico , Anomalía de Pelger-Huët/diagnóstico , Agammaglobulinemia/sangre , Agammaglobulinemia/fisiopatología , Cutis Laxo/diagnóstico , Cutis Laxo/genética , Cutis Laxo/patología , Diagnóstico Diferencial , Tejido Elástico/ultraestructura , Trastornos del Crecimiento/genética , Trastornos del Crecimiento/patología , Humanos , Lactante , Hígado/enzimología , Hígado/patología , Masculino , Enfermedades del Nervio Óptico/genética , Enfermedades del Nervio Óptico/patología , Anomalía de Pelger-Huët/genética , Anomalía de Pelger-Huët/patología , Progeria/diagnóstico , Progeria/genética , Piel/patología , Síndrome , Secuenciación del Exoma , Adulto JovenRESUMEN
Abstract Purpose To evaluate changes in the quantity of elastic fibers in the corpora cavernosa of rats during the natural aging process, and to assess the degree of this change by determining volumetric density (Vv) at different ages via stereological analysis. Methods Forty-eight rats, raised under similar conditions, were subjected to the natural aging process and divided into four groups (G1 to G4), according to age at the time of penectomy (6, 9, 12, and 24 months, respectively). Histological sections of the middle segment of the penis were stained with Weigert's resorcin-fuchsin, and the volumetric density (Vv) of elastic fibers of the corpora cavernosa were determined via stereological analysis. Results There were no statistically significant differences in Vv among groups G1, G2, and G3. These three groups were therefore considered as a single group. The mean Vv of this group showed a statistically significant reduction compared to that of G4 (0.16 vs. 0.11, p<0.05). Conclusion Natural aging in rats was responsible for a reduction in volumetric density of elastic fibers of the corpora cavernosa (approximately 30% decrease in Vv) during senescence.
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Animales , Masculino , Ratas , Pene/citología , Envejecimiento/fisiología , Células Endoteliales/fisiología , Tejido Elástico/ultraestructura , Pene/fisiología , Envejecimiento/patología , Colágeno/fisiología , Colágeno/ultraestructura , Ratas Wistar , Modelos Animales , Tejido Elástico/fisiología , Tejido Elástico/patología , Disfunción Eréctil/fisiopatologíaRESUMEN
BACKGROUND: The introduction of minimally invasive procedures has increased acceptance due to their efficacy, safety, and relatively long-lasting and natural results. At least two different and unrelated modalities are employed to consider a combination treatment. OBJECTIVES: This study aims to evaluate and compare the use and effectiveness of combined microneedling with platelet-rich plasma (PRP) or trichloroacetic acid (TCA) peeling for facial rejuvenation. PATIENTS/METHODS: Twenty-four volunteers with photoaging were randomly divided into three equal groups according to performed procedure on each side of the face (microneedling by dermaroller alone or combined with PRP or TCA 15% peeling). They had received one session every 2 weeks for six sessions of treatment. Photography and punch biopsies were performed before and after 3 months of treatment for clinical, histometrical, and histological evaluation. RESULTS: Combined treatment of dermaroller and PRP or dermaroller and TCA showed significant improvement when compared with dermaroller alone. Significant increase in epidermal thickness was apparent in studied groups, especially after combined treatment with TCA. Organized collagen bundles with newly formed collagen formation and markedly decreased abnormal elastic fibers were noticed in the three studied groups. However, improvement of dermal structures was better demonstrated after combined treatment of Dermaroller and PRP than Dermaroller and TCA 15%. CONCLUSIONS: Most volunteers showed significant clinical improvement after treatment supported by the histometrical and histochemical evaluation; however, the combined use of dermaroller with PRP is apparently more beneficial for facial rejuvenation.
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Técnicas Cosméticas , Plasma Rico en Plaquetas , Rejuvenecimiento , Envejecimiento de la Piel , Cáusticos/uso terapéutico , Quimioexfoliación/efectos adversos , Colágeno/ultraestructura , Terapia Combinada , Técnicas Cosméticas/efectos adversos , Tejido Elástico/ultraestructura , Epidermis/anatomía & histología , Cara , Femenino , Humanos , Persona de Mediana Edad , Agujas , Ácido Tricloroacético/uso terapéuticoRESUMEN
The aim of our study was to assess the value of Elastic stain in the diagnosis of venous invasion (VI) in colonic adenocarcinoma. MATERIAL AND METHODS: All patients who undergone surgery for colonic adenocarcinoma at the University Hospital of Amiens, between 2004 and 2007, were included. Hematein-phloxin-saffron (HPS) stained slides of colectomy specimens were reviewed by two pathologists. Tumor blocks were stained with Elastic Stain (Roche - Ventana®). The presence or absence of VI, their number and localization were correlated with overall survival. RESULTS: Two hundred and thirty-one cases were investigated and 3274 slides were examined. VI were more often diagnosed by Elastic Stain than HPS stain (66% vs. 40%). Ninety percent of VI were revealed within the first 6 HPS slides, and from the first 5 in Elastic Stain. The presence of VI revealed by Elastic Stain and/or HPS was significantly associated with decreased overall survival in multivariate analysis (P=0.029), especially for stage IIA tumors (P=0.016). Tumor differentiation (P=0.006) and pTNM stage (P=0.001) were also independent prognostic factors. The localization and the number of VI were not prognostic factors. CONCLUSION: Our study confirms the prognostic value of VI, revealed by an elastic stain, in colonic adenocarcinoma. A systematic elastic stain of all tumor blocks (number at least 5) could be considered in the future, during pathological examination of colectomy for adenocarcinoma.