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PubMed1997

Galactans and cellulose in flax fibres: putative contributions to the tensile strength.

The proton spin-spin relaxation time, T2, measured from solid-state NMR, indicates a greater rigidity for cellulose than for the adhesive matrix between the microfibrils of flax ultimate fibres. Cytochemical and biochemical analyses allow the identification of: (1) EDTA-soluble RG I-polymers in the primary walls and cell junctions of fibres; (2) long 1 --> 4-beta-D-galactan chains between primary and secondary wall layers; and (3) arabinogalactan-proteins throughout the secondary walls. These polymers in the adhesive matrix between microfibrils and/or cellulose layers ensure that cracks propagate along the matrix rather than across the fibres and play an important role in allowing flax fibres to approach the tensile strength of advanced synthetic fibres like carbon and Kevlar.

Biopolymers↗
PubMed1983

Elastodysplasia and elastodystrophy as the pathologic bases of ocular pterygia and pinguecula.

Specimens of normal conjunctiva from three adult patients were examined by electron microscopy, which revealed normal elastogenesis in the substantia propria, but more prominently, in the episcleral tissues. Ultrastructural examination of eight pterygia and three pinguecula also disclosed evidence of elastogenesis, but in these lesions the morphogenetic sequence of fiber formation was distorted, and the elastic fibers were abnormal. The zone of hyalinization of the substantia propria immediately beneath the epithelium was the only site that manifested clear-cut evidence of collagen degeneration, which assumed the forms of effacement of the longitudinal periodicity of the collagen fibers, and of microfibrillar unfurling of the ends of the collagen fibers. Fibroblastic activity and elastic fiber formation were inconspicuous in this region. Beneath the hyalinized zone were collections of eosinophilic granular material. This material was shown ultrastructurally to be composed of excessive numbers of hollow-centered microfibrils (an elastic fiber precursor), with a tendency to clump centrally in the larger aggregated sheets and to acquire electron-dense inclusions. Numerous fibroblasts were found within this material. Finally, the elastotic fibers of light microscopy represented an abnormal maturational phase of elastic fiber production. These abnormal elastic fibers had microfibrils at their peripheries, but numerous electron-dense inclusions were associated with focal zones of amorphous elastin deposition. Occasionally histiocytes appeared to be engulfing these abnormal fibers. We have concluded that a large component of pinguecula and pterygia is the result of newly synthesized elastic fiber precursors and abnormal maturational forms of elastic fibers (elastodysplasia) that undergo secondary degeneration (elastodystrophy). These structures are presumed to be elaborated by actinically damaged fibroblasts of the substantia propria.

Adult↗
PubMed1995

Elastosis of the lamina cribrosa in pseudoexfoliation syndrome with glaucoma.

BACKGROUND: Pseudoexfoliation syndrome is characterized by the presence of glycoprotein fibers in ocular and extraocular tissues, and often is associated with glaucoma. Pseudoexfoliation material may be associated closely with elastic microfibrillar-associated glycoprotein as well as elastin. METHODS: Four optic nerve heads of two patients with pseudoexfoliation syndrome and glaucoma were examined using electron microscopy and immunogold detection of elastin. Optic nerve heads from healthy age-matched individuals and patients with primary open-angle glaucoma were used for comparisons. RESULTS: In all eyes with pseudoexfoliation and glaucoma, there was marked and widespread elastosis in the connective tissue of the lamina cribrosa. Elastotic fibers appeared as large and irregular aggregates of electron-dense material labeled with anti-elastin antibody. Abundant microfibrils were interspersed in the elastotic aggregates, whereas no typical pseudoexfoliation fibers were observed. In contrast, there were less elastotic fibers in the lamina cribrosa from patients with primary open-angle glaucoma compared with pseudoexfoliation glaucoma. Other changes of extracellular matrix were similar to those observed in primary open-angle glaucoma: decreases in collagen fiber density, presence of basement membranes not associated with cell surfaces, and abundant bundles of microfibrils not labeled with elastin antibody. The elastic fibers appeared normal in other locations within the optic nerves of patients with pseudoexfoliation glaucoma, including in the pial septa and blood vessels of the retrolaminar myelinated optic nerve. CONCLUSION: The authors' findings demonstrate marked and site-specific elastosis in the lamina cribrosa of patients with pseudoexfoliation syndrome with glaucoma, suggesting an abnormal regulation of elastin synthesis and/or degradation in the optic nerve of patients with this disease.

Aged↗
PubMed1988

Schwann cells and collagen synthesis in taxol-treated nerve crush. An electron microscopic study.

The effect of nerve crush on collagen synthesis in rat sciatic nerve was studied by electron microscope. The crushed nerves were treated with taxol which is known to increase the amount of cytoplasmic microtubules at the expense of other cell organelles such as rough endoplasmic reticulum and Golgi complexes. The results were compared to those seen in crushed nerves without taxol treatment. After the injury the amount of collagen fibrils increased at the site of the trauma in both groups when compared to intact controls. Thin (30 mn in diameter) collagen fibrils were often arranged closely to the Schwann cell surface and were connected to deep invaginations in areas where the basal lamina had lost its typical integrity. This was concluded to indicate a probable site of collagen secretion and it provides further evidence that an adult injured nerve Schwann cell is capable of synthesizing fibrous collagen. In taxol-treated nerves additional, abnormally close connection between thin microfibrils of about 10 nm and thin 20-30 nm collagen fibrils appeared in an end-to-end fashion. The microfibrils showed occasional collagenous transverse band like structures. The rough endoplasmic reticulum and Golgi complex play an important role in the posttranslational modifications of the procollagen molecule. Taxol-induced degeneration of cell organelles such as the Golgi complex, which is also essential in the secretion of proteins may thus lead to defective maturation of collagen and may explain partly the altered collagen fibril formation.

Alkaloids↗
PubMed2002

Direct observation of polyhydroxyalkanoate chains by atomic force microscopy.

Atomic force microscopy in the tapping mode was used to investigate aqueous acetone-treated polyhydroxyalkanoate (PHA) inclusions freshly isolated from a recombinant bacterium. The PHA is a copolymer containing about 95 mol% 3-hydroxybutyrate units while the rests are units of 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, and 3-hydroxydodecanoate. Polymer chains extending to several micrometers in length were observed on glass cover slips upon the evaporation of the aqueous acetone. The polymer chains seem to exist in the form of fibrillar aggregates. The height of the microfibrils was about 1 nm. Upon prolonged standing at ambient conditions, the microfibrils dissociated into finer strands of about 0.5 nm in height. The results suggest that biosynthesized PHA are stored in the inclusions in an amorphous state but with minimal chain entanglement. This is possible because the PHA chains exist in the form of fibrillar aggregates that may be the product of a special biosynthesis mechanism.

Cupriavidus necator↗
PubMed2001

DL-penicillamine induced alteration of elastic fibers of periosteum-perichondrium and associated growth inhibition: an experimental study.

Perichondrium-periosteum, being of collagen and elastic fiber, is regarded as a bone growth regulating factor. The aim of the present study was to investigate the role of collagen and elastic fibers on bone growth, by interfering with the fiber assembly in growing chicks upon administration of DL-penicillamine (DL-PNA). Our findings demonstrated that DL-PNA determined relevant modifications in the perichondrium-periosteum, as shown by histochemical, histomorphometrical,biochemical and ultrastructural analysis. This chemical has been shown to inhibit the formation of desmosine cross-links in elastin and to induce an increase of elastin associated microfibrils. On the contrary, the collagen network and the biochemical collagen markers were not affected. These changes resulted in a dramatically reduced growth of long bones in comparison with control. Perichondrial-periosteal regulation of bone growth may be mediated by mechanical and biological factors. This study demonstrates a microstructural change in the perichondrium-periosteum with decreased elastin and increased elastic microfibrils content in penicillamine treated chicks. The mechanism linking changes in the perichondrium-periosteum with altered growth still needs to be elucidated.

Animals↗
PubMed1997

Molecular cloning of the complementary DNA for an additional member of the family of aortic aneurysm antigenic proteins.

PURPOSE: We have purified and partially sequenced a protein from the adventitia of the human aorta (aortic aneurysm antigenic protein 40 kDa; AAAP-40) that has homologies to bovine aortic microfibril-associated glycoprotein (MAGP-36). It is immunoreactive with immunoglobulin G (IgGs) purified from the serum and aortic wall of patients with abdominal aortic aneurysms. AAAP-40 and MAGP-36 have fibrinogen-like and vitronectin-like motifs. Screening an expression library constructed from human aortic adventitial messenger RNA has resulted in the cloning of three complementary DNAs whose gene products are immunoreactive with immunoglobulin G from patients with abdominal aortic aneurysms. Two strongly resemble each other and have been described separately. The purpose of this article is to report the third clone. METHODS: Messenger RNA from a specimen of human aortic aneurysmal adventitia was reverse-transcribed for insertion into the phagemid Uni Zap XR (Stratagene). A strain of Escherichia coli, engineered for expression (XL 1-Blue MFR', Stratagene), was transfected, and rabbit antihuman vitronectin antibody as used to identify positive clones. Sequencing of the positive clones was performed by the Core Laboratories at Columbia University. RESULTS: The hypothetical protein of rAAAP-CL4 (clone 4) shares sequence motifs with known microfibril-associated glycoproteins (MAGPs). The recombinant protein (rAAAP-CL4) is immunoreactive with serum from patients (three of four abdominal aortic aneurysm sera). In addition, similarities have been detected with immunoglobulins of the kappa family and with a protein from cytomegalovirus that is a potential molecular mimic. CONCLUSIONS: There may several members of a novel family of human aortic autoantigenic proteins implicated in abdominal aortic aneurysm disease.

Amino Acid Sequence↗
PubMed2003

Histological study of deposited cementum in human deciduous teeth with pathological root resorption.

Physiological root resorption is a characteristic feature of human deciduous teeth. Pathological root resorption due to apical periodontitis, dental trauma or excessive orthodontic force is also observed in deciduous roots. The root resorption is not continuous, and has resting periods. In the resting period, cementum deposits in resorbed root surface. The deposited cementum in permanent teeth has been reported in detail. However, the deposited cementum in deciduous teeth is unclear. The present study examined apices of roots of human deciduous incisors with apical periodontitis and roots of sound deciduous incisors by light and transmission electron microscopy. Root dentin and original cementum had a severe irregular caved surface. Cementum was partially deposited on the caved root surfaces. The deposited cementum had made the caved root surface relatively flat. The cementum was lax and had some defects. The deposited cementum was belt-like in shape and had a stratified structure. Each layer had various structures consisting of abundant microfibrils and fine granular materials, microfibrils, granular materials, and collagen fibrils, a few fibrils and granular materials and a relatively homogeneous structure. The original cementum had many collagen fibrils, such as intrinsic and extrinsic fibers, and no granular materials or homogeneous structure. Therefore, structure of the deposited cementum was very different from that of original cementum in deciduous teeth and from that of deposited cementum in permanent teeth.

Child↗
PubMed1995

The elastic system of a pressure-bearing tendon of the bullfrog Rana catesbeiana.

Elastic system components have been described in the pressure-bearing tendon of the bullfrog, Rana catesbeiana, as a result of histochemical tests and transmission and scanning electron microscopy. The tension region was shown to possess microfibril bundles, some of which exhibited central deposits of amorphous material. The use of ANS-butanol plus fluorescence microscopy enormously facilitated the identification of elastic system components in both tension and compression regions of the frog tendon. The compression region exhibited pre-elastic and mature elastic fibers, which were shown to be associated with the surface of the convoluted collagen bundles. Thin fibrils were observed in the compression region after ANS treatment. The visceral paratenon had an increased number of elastic fibers located between the collagen bundles and close to the cells. Congo red plus polarization microscopy failed to impart birefringence to the elastic fibers, but they could be identified by their intense staining and isotropic appearance against the bright background of birefringent collagen fibers. SEM demonstrated the three-dimensional aspects of the elastic fibers. They are composed of fibrils of a sinuous nature. The use of ruthenium red in the fixative allowed for the observation of an intimate association of proteoglycan granules with the microfibril bundles. The elastic components identified in the pressure-bearing tendon are assumed to be important for the tissue supramolecular organization, especially in the maintenance of the convoluted state of the collagen fibers in the compression region and their crimp morphology in the tension region. The elastic system must also play an important role in the restoration of the resting shape of the tendon after the deformation achieved during mechanical stimulation.

Animals↗
PubMed2000

Developmental expression of dermatan sulfate proteoglycans in the elastic bovine nuchal ligament.

The nuchal ligament of bovines is a useful system in which to study elastic fibre formation since it contains up to 83% elastin and undergoes a period of rapid elastinogenesis during the last trimester of fetal development and in the first four post-natal months. To identify proteoglycans (PGs) which may be involved in this process we initially investigated changes in the glycosaminoglycan (GAG) profiles during nuchal ligament development. In contrast to the collagenous Achilles tendon, nuchal ligament exhibited: (a) elevated hyaluronan (HA) levels in the peak period of elastin-associated microfibril (fibrillin) synthesis (130-200 days) which precedes elastinogenesis; and (b) markedly increased synthesis of a glucuronate-rich copolymeric form of dermatan sulfate (DS) in the period corresponding to elastin formation (200-270 days). Analysis of DSPGs isolated from 230-day nuchal ligament showed that this copolymer was predominantly associated with a glycoform of biglycan which was specifically elevated at this stage in development. This finding was consistent with Northern blot analysis which showed that steady-state biglycan mRNA levels increased significantly during the elastinogenic period. In contrast, the mRNA levels for decorin, the only other DSPG detected in this tissue, declined rapidly after 140 days of fetal development. In conclusion, the results suggest that HA may play a role in microfibril assembly and that a specific glycoform of biglycan may be associated with the elastinogenic phase of elastic fibre formation.

Animals↗
PubMed2001

Synthesis and structural organization of zonular fibers during development and aging.

Zonular fibers are a specific form of extracellular matrix composed mainly of fibrillins. The purpose of this study was to determine which cells secrete fibrillin-1 during development and aging. A specific guinea pig fibrillin-1 mRNA probe was designed and cloned in order to identify fibrillin-secreting cells in guinea pig eye, using in situ hybridization. Immunofluorescence, with a specific guinea pig monoclonal antibody, was used to compare protein levels at different stages from birth to 35 months of age. Electron microscopy and immunolabeling were used to investigate the organization of zonular microfibril bundles. We identified the cells of non-pigmented epithelium of the ciliary body as the main source of fibrillin secreted into the zonule. Moreover, while mRNA expression decreased during aging, there was no decrease in fibrillin immunoreactivity, as previously described in human aorta. These data indicate a very slow turnover of the zonular microfibrils which can be correlated with the appearance during aging of a new periodic fibrillar structure. This new structure may reflect an increased cross-linking in the long-lived zonular microfibrillar bundles.

Aging↗
PubMed1996

Fibrillln mutations in Marfan syndrome and related phenotypes.

A casual association has been established between mutations in the fibrillin 1 gene and Marfan syndrome and related phenotypes. Analysis of mutations in these disease types has provided new insights into microfibril assembly and function. These include evidence for a mutation in a fibrillin 1 domain associated with severe phenotype; indication of profibrillin processing by a furin-like endoprotease; linkage between extracellular processing and fibrillin 1 polymerization; and involvement of calcium binding in monomer stabilization and microfibril assembly. Identification of intragenic DNA polymorphisms and determination of intron/exon junction sequences have significantly improved our ability to diagnose Marfan syndrome and to detect fibrillin 1 mutations. Additional work has provided strong evidence for structural and functional heterogeneity of microfibrillin. The evidence includes the identification of fibrillin 2, a microfibrillar component structurally related to fibrillin 1; the differential pattern of gene expression of the two fibrillin; and the association of fibrillin 2 mutations with congenital contractural arachnodactyly.

Animals↗
PubMed1996

The Marfan syndrome and the cardiovascular surgeon.

The authors present the current status of surgery for the cardiovascular manifestations of the Marfan syndrome. In addition, a brief review of current Marfan genetic research is presented. Data on all Marfan patients undergoing aortic root replacement at the Johns Hopkins Hospital (September 1976-June 1995) were analyzed. Survival and event-free curves were calculated and risk factors for early and late death were determined by univariate and multivariate analysis. Two hundred twelve Marfan patients underwent aortic root replacement using composite graft (202), homograft (8) or valve-sparing procedures (2). One hundred eighty-five patients underwent elective repair with no 30-day mortality. Twenty-seven patients underwent urgent surgery, primarily for acute dissection; two patients with aortic rupture died in the operating room. Actuarial survival of the 212 patients was 88% at 5 years, 78% at 10 years and 71% at 14 years. By multivariate analysis, only poor NYHA class, male gender and urgent surgery emerged as significant independent predictors of early or late mortality. Histologic examination of excised Marfan aortic leaflets by immunofluorescent staining for fibrillin showed fragmentation of elastin-associated microfibrils. These studies suggest cautious use of valve-sparing procedures in Marfan patients. Over the last 5 years significant progress has been made in identifying mutant genes that code for defective fibrillin microfibrils in Marfan patients. Attempts are underway to develop animal models of Marfan disease for study of possible gene therapy. Aortic root replacement can be performed in Marfan patients with operative risk under 5%. Long-term results are gratifying. At present, valve-sparing procedures should be used cautiously in Marfan patients because of fibrillin abnormalities in the preserved aortic valve leaflets.

Adolescent↗
PubMed1999

Latent transforming growth factor-beta binding proteins (LTBPs)--structural extracellular matrix proteins for targeting TGF-beta action.

Growth factors of the transforming growth factor-beta family are potent regulators of the extracellular matrix formation, in addition to their immunomodulatory and regulatory roles for cell growth. TGF-beta s are secreted from cells as latent complexes containing TGF-beta and its propeptide, LAP (latency-associated peptide). In most cells LAP is covalently linked to an additional protein, latent TGF-beta binding protein (LTBP), forming the large latent complex. LTBPs are required for efficient secretion and correct folding of TGF-beta s. The secreted large latent complexes associate covalently with the extracellular matrix via the N-termini of the LTBPs. LTBPs belong to the fibrillin-LTBP family of extracellular matrix proteins, which have a typical repeated domain structure consisting mostly of epidermal growth factor (EGF)-like repeats and characteristic eight cysteine (8-Cys) repeats. Currently four different LTBPs and two fibrillins have been identified. LTBPs contain multiple proteinase sensitive sites, providing means to solubilize the large latent complex from the extracellular matrix structures. LTBPs are now known to exist both as soluble molecules and in association with the extracellular matrix. An important consequence of this is LTBP-mediated deposition and targeting of latent, activatable TGF-beta into extracellular matrices and connective tissues. LTBPs have a dual function, they are required both for the secretion of the small latent TGF-beta complex as well as directing bound latent TGF-beta to extracellular matrix microfibrils. However, it is not known at present whether LTBPs are capable of forming microfibrils independently, or whether they are a part of the fibrillin-containing fibrils. Most LTBPs possess RGD-sequences, which may have a role in their interactions with the cell surface. At least LTBP-1 is chemotactic to smooth muscle cells, and is involved in vascular remodelling. Analyses of the expressed LTBPs have revealed considerable variations throughout the molecules, generated both by alternative splicing and utilization of multiple promoter regions. The significance of this structural diversity is mostly unclear at present.

Adaptor Proteins, Signal Transducing↗
PubMed1991

Ultrastructure and microanalyses of the protoscolex hooks of Echinococcus granulosus.

The rostellar distal cytoplasm of Echinococcus granulosus protoscoleces is characterized by extensive basal membrane infolding, prominent hemidesmosomes and is subtended by a lamina reticularis with microfibrils of approximately 10 nm diameter that occasionally show a 55 nm banding periodicity. The rostellar hooks, in 2 rows, each have a blade, guard and handle region and possess a central amorphous pulp, a middle microfibrillar medulla with microfibrils of approximately 4 nm diameter, and a complex outer cortex in all but the proximal region of the guard and the base of the handle. In these regions additional material, of similar electron density to the medulla, but lacking the fibrillar substructure, occurs and gives the areas a lobed appearance. Energy-dispersive X-ray microanalysis of whole hooks demonstrated the presence of sulphur and trace quantities of phosphorus. X-ray near-edge absorption spectra resembled those of cystine, feather and hair and showed the sulphur to be predominantly in the form of disulphide linkages. X-ray diffraction patterns of whole hook preparations revealed 2 diffuse rings with equatorial spacings of 7.99 A and 15.22 A, thus differing from vertebrate keratins.

Animals↗
PubMed2005

Microscopic examination of chitosan-polyphosphate beads with entrapped spores of the biocontrol agent, Streptomyces melanosporofaciens EF-76.

Spores of the biocontrol agent, Streptomyces melanosporofaciens EF-76, were entrapped by complex coacervation in beads composed of a macromolecular complex (MC) of chitosan and polyphosphate. A proportion of spores entrapped in beads survived the entrapment procedure as shown by treating spores from chitosan beads with a dye allowing the differentiation of live and dead cells. The spore-loaded chitosan beads could be digested by a chitosanase, suggesting that, once introduced in soil, the beads would be degraded to release the biocontrol agent. Spore-loaded beads were examined by optical and scanning electron microscopy because the release of the biological agent depends on the spore distribution in the chitosan beads. The microscopic examination revealed that the beads had a porous surface and contained a network of inner microfibrils. Spores were entrapped in both the chitosan microfibrils and the bead lacuna.

Chitosan↗
PubMed2000

Periodate oxidation of crystalline cellulose.

Conversion of 1,2-dihydroxyl groups to dialdehyde by periodate oxidation is a useful method of derivatizing polysaccharides but has not been extensively utilized in derivatization of cellulose because of complicacy due to the crystalline nature of cellulose. To understand the influence of cellulose crystallinity on this reaction, we investigated how the periodate oxidation proceeds with a highly crystalline cellulose of the marine alga Cladophora sp. The crystallinity of the oxidized cellulose, determined by X-ray diffraction, decreased according to the oxidation level. The half-height widths of equatorial diffraction peaks were nearly unchanged. The solid-state 13C NMR spectra did not show peaks corresponding to aldehyde groups, but solution 13C NMR spectra showed the presence of dicarboxylic groups after subsequent oxidation by sodium chlorite. Transmission electron microscopy showed that microfibrils of Cladophora tended to be bent and more flexible than the original sample. Gold labeling of the aldehyde groups, mediated by thiosemicarbazide derivatization, revealed a highly uneven distribution of dialdehyde groups. When treated by 50% (w/v) sulfuric acid, partially oxidized Cladophora cellulose gave many short fragments of microfibril. These features indicate that the periodate oxidation proceeds by forming dialdehyde groups in longitudinally spaced, bandlike domains.

Aldehydes↗
PubMed2001

A small-angle X-ray scattering study of the effect of hydration on the microstructure of flax fibers.

Small-angle X-ray scattering (SAXS) experiments with in situ dehydration have been performed on small bundles of flax fibers. Analysis indicated that the structure of the cellulose microfibrils is consistent with a cross section of approximately 10 x 50 A2. The microfibril misalignment was Gaussian with a full width at half-maximum of 11-15 degrees, dependent on hydration level. A meridional peak at q = 0.095 A-1 was observed, possibly for the first time in cellulose I fibers, and this has been attributed to a crystalline/noncrystalline repeat repeat distance of 60-70 A. It has also been noted that there is considerable change in structure as flax fibers dry from the wet state.

Cell Wall↗