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Sample records for a-type nuclear lamins

  1. A-type Lamins Form Distinct Filamentous Networks with Differential Nuclear Pore Complex Associations.

    PubMed

    Xie, Wei; Chojnowski, Alexandre; Boudier, Thomas; Lim, John S Y; Ahmed, Sohail; Ser, Zheng; Stewart, Colin; Burke, Brian

    2016-10-10

    The nuclear lamina is a universal feature of metazoan nuclear envelopes (NEs) [1]. In mammalian cells, it appears as a 10-30 nm filamentous layer at the nuclear face of the inner nuclear membrane (INM) and is composed primarily of A- and B-type lamins, members of the intermediate filament family [2]. While providing structural integrity to the NE, the lamina also represents an important signaling and regulatory platform [3]. Two A-type lamin isoforms, lamins A and C (LaA and LaC), are expressed in most adult human cells. Encoded by a single gene, these proteins are largely identical, diverging only in their C-terminal tail domains. By contrast with that of LaC, the unique LaA tail undergoes extensive processing, including farnesylation and endo-proteolysis [4, 5]. However, functional differences between LaA and LaC are still unclear. Compounding this uncertainty, the structure of the lamina remains ill defined. In this study, we used BioID, an in vivo proximity-labeling method to identify differential interactors of A-type lamins [6]. One of these, Tpr, a nuclear pore complex (NPC) protein, is highlighted by its selective association with LaC. By employing superresolution microscopy, we demonstrate that this Tpr association is mirrored in enhanced interaction of LaC with NPCs. Further superresolution studies visualizing both endogenous A- and B-type lamins have allowed us to construct a nanometer-scale model of the mammalian nuclear lamina. Our data indicate that different A- and B-type lamin species assemble into separate filament networks that together form an extended composite structure at the nuclear periphery providing attachment sites for NPCs, thereby regulating their distribution.

  2. Proteasome-mediated degradation of integral inner nuclear membrane protein emerin in fibroblasts lacking A-type lamins

    SciTech Connect

    Muchir, Antoine; Massart, Catherine; Engelen, Baziel G. van; Lammens, Martin; Bonne, Gisele; Worman, Howard J. . E-mail: hjw14@columbia.edu

    2006-12-29

    We previously identified and characterized a homozygous LMNA nonsense mutation leading to the absence of A-type lamins in a premature neonate who died at birth. We show here that the absence of A-type lamins is due to degradation of the aberrant mRNA transcript with a premature termination codon. In cultured fibroblasts from the subject with the homozygous LMNA nonsense mutation, there was a decreased steady-state expression of the integral inner nuclear membrane proteins emerin and nesprin-1{alpha} associated with their mislocalization to the bulk endoplasmic reticulum and a hyperphosphorylation of emerin. To determine if decreased emerin expression occurred post-translationally, we treated cells with a selective proteasome inhibitor and observed an increase in expression. Our results show that mislocalization of integral inner nuclear membrane proteins to the endoplasmic reticulum in human cells lacking A-type lamins leads to their degradation and provides the first evidence that their degradation is mediated by the proteasome.

  3. A-type and B-type lamins initiate layer assembly at distinct areas of the nuclear envelope in living cells

    SciTech Connect

    Furukawa, Kazuhiro; Ishida, Kazuya; Tsunoyama, Taka-aki; Toda, Suguru; Osoda, Shinichi; Horigome, Tsuneyoshi; Fisher, Paul A.; Sugiyama, Shin

    2009-04-15

    To investigate nuclear lamina re-assembly in vivo, Drosophila A-type and B-type lamins were artificially expressed in Drosophila lamin Dm{sub 0}null mutant brain cells. Both exogenous lamin C (A-type) and Dm{sub 0} (B-type) formed sub-layers at the nuclear periphery, and efficiently reverted the abnormal clustering of the NPC. Lamin C initially appeared where NPCs were clustered, and subsequently extended along the nuclear periphery accompanied by the recovery of the regular distribution of NPCs. In contrast, lamin Dm{sub 0} did not show association with the clustered NPCs during lamina formation and NPC spacing recovered only after completion of a closed lamin Dm{sub 0} layer. Further, when lamin Dm{sub 0} and C were both expressed, they did not co-polymerize, initiating layer formation in separate regions. Thus, A and B-type lamins reveal differing properties during lamina assembly, with A-type having the primary role in organizing NPC distribution. This previously unknown complexity in the assembly of the nuclear lamina could be the basis for intricate nuclear envelope functions.

  4. Loss of Drosophila A-type lamin C initially causes tendon abnormality including disintegration of cytoskeleton and nuclear lamina in muscular defects.

    PubMed

    Uchino, Ryo; Nonaka, Yu-Ki; Horigome, Tuneyoshi; Sugiyama, Shin; Furukawa, Kazuhiro

    2013-01-01

    Lamins are the major components of nuclear envelope architecture, being required for both the structural and informational roles of the nuclei. Mutations of lamins cause a spectrum of diseases in humans, including muscular dystrophy. We report here that the loss of the A-type lamin gene, lamin C in Drosophila resulted in pupal metamorphic lethality caused by tendon defects, matching the characteristics of human A-type lamin revealed by Emery-Dreifuss muscular dystrophy (EDMD). In tendon cells lacking lamin C activity, overall cell morphology was affected and organization of the spectraplakin family cytoskeletal protein Shortstop which is prominently expressed in tendon cells gradually disintegrated, notably around the nucleus and in a manner correlating well with the degradation of musculature. Furthermore, lamin C null mutants were efficiently rescued by restoring lamin C expression to shortstop-expressing cells, which include tendon cells but exclude skeletal muscle cells. Thus the critical function of A-type lamin C proteins in Drosophila musculature is to maintain proper function and morphology of tendon cells.

  5. Nuclear lamins and neurobiology.

    PubMed

    Young, Stephen G; Jung, Hea-Jin; Lee, John M; Fong, Loren G

    2014-08-01

    Much of the work on nuclear lamins during the past 15 years has focused on mutations in LMNA (the gene for prelamin A and lamin C) that cause particular muscular dystrophy, cardiomyopathy, partial lipodystrophy, and progeroid syndromes. These disorders, often called "laminopathies," mainly affect mesenchymal tissues (e.g., striated muscle, bone, and fibrous tissue). Recently, however, a series of papers have identified important roles for nuclear lamins in the central nervous system. Studies of knockout mice uncovered a key role for B-type lamins (lamins B1 and B2) in neuronal migration in the developing brain. Also, duplications of LMNB1 (the gene for lamin B1) have been shown to cause autosome-dominant leukodystrophy. Finally, recent studies have uncovered a peculiar pattern of nuclear lamin expression in the brain. Lamin C transcripts are present at high levels in the brain, but prelamin A expression levels are very low-due to regulation of prelamin A transcripts by microRNA 9. This form of prelamin A regulation likely explains why "prelamin A diseases" such as Hutchinson-Gilford progeria syndrome spare the central nervous system. In this review, we summarize recent progress in elucidating links between nuclear lamins and neurobiology.

  6. Nuclear Lamins: Thin Filaments with Major Functions.

    PubMed

    de Leeuw, Rebecca; Gruenbaum, Yosef; Medalia, Ohad

    2017-09-08

    The nuclear lamina is a nuclear peripheral meshwork that is mainly composed of nuclear lamins, although a small fraction of lamins also localizes throughout the nucleoplasm. Lamins are classified as type V intermediate filament (IF) proteins. Mutations in lamin genes cause at least 15 distinct human diseases, collectively termed laminopathies, including muscle, metabolic, and neuronal diseases, and can cause accelerated aging. Most of these mutations are in the LMNA gene encoding A-type lamins. A growing number of nuclear proteins are known to bind lamins and are implicated in both nuclear and cytoskeletal organization, mechanical stability, chromatin organization, signaling, gene regulation, genome stability, and cell differentiation. Recent studies reveal the organization of the lamin filament meshwork in somatic cells where they assemble as tetramers in cross-section of the filaments. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Both lamin A and lamin C mutations cause lamina instability as well as loss of internal nuclear lamin organization

    SciTech Connect

    Broers, Jos L.V. . E-mail: jos.broers@molcelb.unimaas.nl; Kuijpers, H.J.H.; Oestlund, C.; Worman, H.J.; Endert, J.; Ramaekers, F.C.S.

    2005-04-01

    We have applied the fluorescence loss of intensity after photobleaching (FLIP) technique to study the molecular dynamics and organization of nuclear lamin proteins in cell lines stably transfected with green fluorescent protein (GFP)-tagged A-type lamin cDNA. Normal lamin A and C proteins show abundant decoration of the inner layer of the nuclear membrane, the nuclear lamina, and a generally diffuse localization in the nuclear interior. Bleaching studies revealed that, while the GFP-tagged lamins in the lamina were virtually immobile, the intranuclear fraction of these molecules was partially mobile. Intranuclear lamin C was significantly more mobile than intranuclear lamina A. In search of a structural cause for the variety of inherited diseases caused by A-type lamin mutations, we have studied the molecular organization of GFP-tagged lamin A and lamin C mutants R453W and R386K, found in Emery-Dreifuss muscular dystrophy (EDMD), and lamin A and lamin C mutant R482W, found in patients with Dunnigan-type familial partial lipodystrophy (FPLD). In all mutants, a prominent increase in lamin mobility was observed, indicating loss of structural stability of lamin polymers, both at the perinuclear lamina and in the intranuclear lamin organization. While the lamin rod domain mutant showed overall increased mobility, the tail domain mutants showed mainly intranuclear destabilization, possibly as a result of loss of interaction with chromatin. Decreased stability of lamin mutant polymers was confirmed by flow cytometric analyses and immunoblotting of nuclear extracts. Our findings suggest a loss of function of A-type lamin mutant proteins in the organization of intranuclear chromatin and predict the loss of gene regulatory function in laminopathies.

  8. Mechanical model of blebbing in nuclear lamin meshworks.

    PubMed

    Funkhouser, Chloe M; Sknepnek, Rastko; Shimi, Takeshi; Goldman, Anne E; Goldman, Robert D; Olvera de la Cruz, Monica

    2013-02-26

    Much of the structural stability of the nucleus comes from meshworks of intermediate filament proteins known as lamins forming the inner layer of the nuclear envelope called the nuclear lamina. These lamin meshworks additionally play a role in gene expression. Abnormalities in nuclear shape are associated with a variety of pathologies, including some forms of cancer and Hutchinson-Gilford Progeria Syndrome, and often include protruding structures termed nuclear blebs. These nuclear blebs are thought to be related to pathological gene expression; however, little is known about how and why blebs form. We have developed a minimal continuum elastic model of a lamin meshwork that we use to investigate which aspects of the meshwork could be responsible for bleb formation. Mammalian lamin meshworks consist of two types of lamin proteins, A type and B type, and it has been reported that nuclear blebs are enriched in A-type lamins. Our model treats each lamin type separately and thus, can assign them different properties. Nuclear blebs have been reported to be located in regions where the fibers in the lamin meshwork have a greater separation, and we find that this greater separation of fibers is an essential characteristic for generating nuclear blebs. The model produces structures with comparable morphologies and distributions of lamin types as real pathological nuclei. Thus, preventing this opening of the meshwork could be a route to prevent bleb formation, which could be used as a potential therapy for the pathologies associated with nuclear blebs.

  9. An Elastic Model of Blebbing in Nuclear Lamin Meshworks

    NASA Astrophysics Data System (ADS)

    Funkhouser, Chloe; Sknepnek, Rastko; Shimi, Takeshi; Goldman, Anne; Goldman, Robert; Olvera de La Cruz, Monica

    2013-03-01

    A two-component continuum elastic model is introduced to analyze a nuclear lamin meshwork, a structural element of the lamina of the nuclear envelope. The main component of the lamina is a meshwork of lamin protein filaments providing mechanical support to the nucleus and also playing a role in gene expression. Abnormalities in nuclear shape are associated with a variety of pathologies, including some forms of cancer and Hutchinson-Gilford progeria syndrome, and are often characterized by protruding structures termed nuclear blebs. Nuclear blebs are rich in A-type lamins and may be related to pathological gene expression. We apply the two-dimensional elastic shell model to determine which characteristics of the meshwork could be responsible for blebbing, including heterogeneities in the meshwork thickness and mesh size. We find that if one component of the lamin meshwork, rich in A-type lamins, has a tendency to form a larger mesh size than that rich in B-type lamins, this is sufficient to cause segregation of the lamin components and also to form blebs rich in A-type lamins. The model produces structures with comparable morphologies and mesh size distributions as the lamin meshworks of real, pathological nuclei. Funded by US DoE Award DEFG02-08ER46539 and by the DDR&E and AFOSR under Award FA9550-10-1-0167; simulations performed on NU Quest cluster

  10. A Comparative Study of Drosophila and Human A-Type Lamins

    PubMed Central

    Schulze, Sandra R.; Curio-Penny, Beatrice; Speese, Sean; Dialynas, George; Cryderman, Diane E.; McDonough, Caitrin W.; Nalbant, Demet; Petersen, Melissa; Budnik, Vivian; Geyer, Pamela K.; Wallrath, Lori L.

    2009-01-01

    Nuclear intermediate filament proteins, called lamins, form a meshwork that lines the inner surface of the nuclear envelope. Lamins contain three domains: an N-terminal head, a central rod and a C-terminal tail domain possessing an Ig-fold structural motif. Lamins are classified as either A- or B-type based on structure and expression pattern. The Drosophila genome possesses two genes encoding lamins, Lamin C and lamin Dm0, which have been designated A- and B-type, respectively, based on their expression profile and structural features. In humans, mutations in the gene encoding A-type lamins are associated with a spectrum of predominantly tissue-specific diseases known as laminopathies. Linking the disease phenotypes to cellular functions of lamins has been a major challenge. Drosophila is being used as a model system to identify the roles of lamins in development. Towards this end, we performed a comparative study of Drosophila and human A-type lamins. Analysis of transgenic flies showed that human lamins localize predictably within the Drosophila nucleus. Consistent with this finding, yeast two-hybrid data demonstrated conservation of partner-protein interactions. Drosophila lacking A-type lamin show nuclear envelope defects similar to those observed with human laminopathies. Expression of mutant forms of the A-type Drosophila lamin modeled after human disease-causing amino acid substitutions revealed an essential role for the N-terminal head and the Ig-fold in larval muscle tissue. This tissue-restricted sensitivity suggests a conserved role for lamins in muscle biology. In conclusion, we show that (1) localization of A-type lamins and protein-partner interactions are conserved between Drosophila and humans, (2) loss of the Drosophila A-type lamin causes nuclear defects and (3) muscle tissue is sensitive to the expression of mutant forms of A-type lamin modeled after those causing disease in humans. These studies provide new insights on the role of lamins in

  11. Tissue specific loss of A-type lamins in the gastrointestinal epithelium can enhance polyp size.

    PubMed

    Wang, Audrey S; Kozlov, Serguei V; Stewart, Colin L; Horn, Henning F

    2015-01-01

    The nuclear lamina, comprised of the A and B-type lamins, is important in maintaining nuclear shape and in regulating key nuclear functions such as chromatin organization and transcription. Deletion of the A-type lamins results in genome instability and many cancers show altered levels of A-type lamin expression. Loss of function mutations in the mouse Lmna gene result in early postnatal lethality, usually within 3-5 weeks of birth making an analysis of the role of lamins in carcinogenesis difficult. To circumvent early lethality, and determine the role of the A-type lamins in specific tissues in older mice we derived a conditional allele of Lmna(FL/FL) (floxed). Lmna(FL/FL) was specifically deleted in the gastrointestinal (GI) epithelium by crossing the Lmna(FL/FL) mice with Villin-Cre mice. Mice lacking Lmna in the GI are overtly normal with no effects on overall growth, longevity or GI morphology. On a GI specific sensitized (Apc(Min/+)) background, polyp numbers are unchanged, but polyp size is slightly increased, and only in the duodenum. Our findings reveal that although A-type lamins are dispensable in the postnatal GI epithelium, loss of Lmna under malignant conditions may, to a limited extent, enhance polyp size indicating that A-type lamins may regulate cell proliferation in the transformed GI epithelium. Published by Elsevier B.V.

  12. Role of A-type lamins in signaling, transcription, and chromatin organization

    PubMed Central

    González, José M.

    2009-01-01

    A-type lamins (lamins A and C), encoded by the LMNA gene, are major protein constituents of the mammalian nuclear lamina, a complex structure that acts as a scaffold for protein complexes that regulate nuclear structure and functions. Interest in these proteins has increased in recent years with the discovery that LMNA mutations cause a variety of human diseases termed laminopathies, including progeroid syndromes and disorders that primarily affect striated muscle, adipose, bone, and neuronal tissues. In this review, we discuss recent research supporting the concept that lamin A/C and associated nuclear envelope proteins regulate gene expression in health and disease through interplay with signal transduction pathways, transcription factors, and chromatin-associated proteins. PMID:20038676

  13. Nuclear lamins and oxidative stress in cell proliferation and longevity.

    PubMed

    Shimi, Takeshi; Goldman, Robert D

    2014-01-01

    In mammalian cells, the nuclear lamina is composed of a complex fibrillar network associated with the inner membrane of the nuclear envelope. The lamina provides mechanical support for the nucleus and functions as the major determinant of its size and shape. At its innermost aspect it associates with peripheral components of chromatin and thereby contributes to the organization of interphase chromosomes. The A- and B-type lamins are the major structural components of the lamina, and numerous mutations in the A-type lamin gene have been shown to cause many types of human diseases collectively known as the laminopathies. These mutations have also been shown to cause a disruption in the normal interactions between the A and B lamin networks. The impact of these mutations on nuclear functions is related to the roles of lamins in regulating various essential processes including DNA synthesis and damage repair, transcription and the regulation of genes involved in the response to oxidative stress. The major cause of oxidative stress is the production of reactive oxygen species (ROS), which is critically important for cell proliferation and longevity. Moderate increases in ROS act to initiate signaling pathways involved in cell proliferation and differentiation, whereas excessive increases in ROS cause oxidative stress, which in turn induces cell death and/or senescence. In this review, we cover current findings about the role of lamins in regulating cell proliferation and longevity through oxidative stress responses and ROS signaling pathways. We also speculate on the involvement of lamins in tumor cell proliferation through the control of ROS metabolism.

  14. Broken nuclei--lamins, nuclear mechanics, and disease.

    PubMed

    Davidson, Patricia M; Lammerding, Jan

    2014-04-01

    Mutations in lamins, which are ubiquitous nuclear intermediate filaments, lead to a variety of disorders including muscular dystrophy and dilated cardiomyopathy. Lamins provide nuclear stability, help connect the nucleus to the cytoskeleton, and can modulate chromatin organization and gene expression. Nonetheless, the diverse functions of lamins remain incompletely understood. We focus here on the role of lamins on nuclear mechanics and their involvement in human diseases. Recent findings suggest that lamin mutations can decrease nuclear stability, increase nuclear fragility, and disturb mechanotransduction signaling, possibly explaining the muscle-specific defects in many laminopathies. At the same time, altered lamin expression has been reported in many cancers, where the resulting increased nuclear deformability could enhance the ability of cells to transit tight interstitial spaces, thereby promoting metastasis.

  15. Nuclear Envelope Lamin-A Couples Actin Dynamics with Immunological Synapse Architecture and T Cell Activation

    PubMed Central

    González-Granado, José María; Trigueros-Motos, Laia; Cibrián, Danay; Morlino, Giulia; Blanco-Berrocal, Marta; Osorio, Fernando Garcia; Freije, José María Pérez; López-Otín, Carlos; Sánchez-Madrid, Francisco; Andrés, Vicente

    2014-01-01

    In many cell types, nuclear A-type lamins have been implicated in structural and functional activities, including higher-order genome organization, DNA replication and repair, gene transcription, and signal transduction. However, their role in specialized immune cells remains largely unexplored. Here, we showed that the abundance of A-type lamins is almost negligible in resting naïve T lymphocytes, but that it is substantially increased upon activation of the T cell receptor (TCR), and is an early event that accelerates formation of the immunological synapse between T cells and antigen-presenting cells. We found that lamin-A enhanced the polymerization of F-actin in T cells, a critical step for immunological synapse formation, by physically connecting the nucleus to the plasma membrane through the linker of nucleoskeleton and cytoskeleton (LINC) complex. We also showed that lamin-A played a key role in other membrane, cytoplasmic, and nuclear events related to TCR activation, including receptor-clustering, downstream signaling, and target gene expression. Notably, the presence of lamin-A was associated with enhanced extracellular signal–regulated kinase 1/2 signaling, and pharmacological inhibition of this pathway reduced the extent of lamin-A–dependent T cell activation. Moreover, mice deficient in lamin-A exhibited impaired T cell responses in vivo. These findings underscore the importance of A-type lamins for TCR activation, and identify lamin-A as a previously unappreciated regulator of the immune response. PMID:24757177

  16. Nuclear protein import is reduced in cells expressing nuclear envelopathy-causing lamin A mutants

    SciTech Connect

    Busch, Albert; Kiel, Tilman; Heupel, Wolfgang-M.; Wehnert, Manfred; Huebner, Stefan

    2009-08-15

    Lamins, which form the nuclear lamina, not only constitute an important determinant of nuclear architecture, but additionally play essential roles in many nuclear functions. Mutations in A-type lamins cause a wide range of human genetic disorders (laminopathies). The importance of lamin A (LaA) in the spatial arrangement of nuclear pore complexes (NPCs) prompted us to study the role of LaA mutants in nuclear protein transport. Two mutants, causing prenatal skin disease restrictive dermopathy (RD) and the premature aging disease Hutchinson Gilford progeria syndrome, were used for expression in HeLa cells to investigate their impact on the subcellular localization of NPC-associated proteins and nuclear protein import. Furthermore, dynamics of the LaA mutants within the nuclear lamina were studied. We observed affected localization of NPC-associated proteins, diminished lamina dynamics for both LaA mutants and reduced nuclear import of representative cargo molecules. Intriguingly, both LaA mutants displayed similar effects on nuclear morphology and functions, despite their differences in disease severity. Reduced nuclear protein import was also seen in RD fibroblasts and impaired lamina dynamics for the nucleoporin Nup153. Our data thus represent the first study of a direct link between LaA mutant expression and reduced nuclear protein import.

  17. A-type lamins and Hutchinson-Gilford progeria syndrome: pathogenesis and therapy.

    PubMed

    Gonzalez, Jose M; Pla, Davinia; Perez-Sala, Dolores; Andres, Vicente

    2011-06-01

    Lamin A and lamin C (A-type lamins, both encoded by the LMNA gene) are major components of the mammalian nuclear lamina, a complex proteinaceous structure that acts as a scaffold for protein complexes that regulate nuclear structure and function. Abnormal accumulation of farnesylated-progerin, a mutant form of prelamin A, plays a key role in the pathogenesis of the Hutchinson-Gilford progeria syndrome (HGPS), a devastating disorder that causes the death of affected children at an average age of 13.5 years, predominantly from premature atherosclerosis and myocardial infarction or stroke. Remarkably, progerin is also present in normal cells and appears to progressively accumulate during aging of non-HGPS cells. Therefore, understanding how this mutant form of lamin A provokes HGPS may shed significant insight into physiological aging. In this review, we discuss recent advances into the pathogenic mechanisms underlying HGPS, the main murine models of the disease, and the therapeutic strategies developed in cellular and animal models with the aim of reducing the accumulation of farnesylated-progerin, as well as their use in clinical trials of HGPS.

  18. The nucleoporin Nup88 is interacting with nuclear lamin A

    PubMed Central

    Lussi, Yvonne C.; Hügi, Ilona; Laurell, Eva; Kutay, Ulrike; Fahrenkrog, Birthe

    2011-01-01

    Nuclear pore complexes (NPCs) are embedded in the nuclear envelope (NE) and mediate bidirectional nucleocytoplasmic transport. Their spatial distribution in the NE is organized by the nuclear lamina, a meshwork of nuclear intermediate filament proteins. Major constituents of the nuclear lamina are A- and B-type lamins. In this work we show that the nuclear pore protein Nup88 binds lamin A in vitro and in vivo. The interaction is mediated by the N-terminus of Nup88, and Nup88 specifically binds the tail domain of lamin A but not of lamins B1 and B2. Expression of green fluorescent protein–tagged lamin A in cells causes a masking of binding sites for Nup88 antibodies in immunofluorescence assays, supporting the interaction of lamin A with Nup88 in a cellular context. The epitope masking disappears in cells expressing mutants of lamin A that are associated with laminopathic diseases. Consistently, an interaction of Nup88 with these mutants is disrupted in vitro. Immunoelectron microscopy using Xenopus laevis oocyte nuclei further revealed that Nup88 localizes to the cytoplasmic and nuclear face of the NPC. Together our data suggest that a pool of Nup88 on the nuclear side of the NPC provides a novel, unexpected binding site for nuclear lamin A. PMID:21289091

  19. Nuclear envelope lamin-A couples actin dynamics with immunological synapse architecture and T cell activation.

    PubMed

    González-Granado, José M; Silvestre-Roig, Carlos; Rocha-Perugini, Vera; Trigueros-Motos, Laia; Cibrián, Danay; Morlino, Giulia; Blanco-Berrocal, Marta; Osorio, Fernando G; Freije, José M P; López-Otín, Carlos; Sánchez-Madrid, Francisco; Andrés, Vicente

    2014-04-22

    In many cell types, nuclear A-type lamins regulate multiple cellular functions, including higher-order genome organization, DNA replication and repair, gene transcription, and signal transduction; however, their role in specialized immune cells remains largely unexplored. We showed that the abundance of A-type lamins was almost negligible in resting naïve T lymphocytes, but was increased upon activation of the T cell receptor (TCR). The increase in lamin-A was an early event that accelerated formation of the immunological synapse between T cells and antigen-presenting cells. Polymerization of F-actin in T cells is a critical step for immunological synapse formation, and lamin-A interacted with the linker of nucleoskeleton and cytoskeleton (LINC) complex to promote F-actin polymerization. We also showed that lamin-A expression accelerated TCR clustering and led to enhanced downstream signaling, including extracellular signal-regulated kinase 1/2 (ERK1/2) signaling, as well as increased target gene expression. Pharmacological inhibition of the ERK pathway reduced lamin-A-dependent T cell activation. Moreover, mice lacking lamin-A in immune cells exhibited impaired T cell responses in vivo. These findings underscore the importance of A-type lamins for TCR activation and identify lamin-A as a previously unappreciated regulator of the immune response.

  20. Concentration-dependent Effects of Nuclear Lamins on Nuclear Size in Xenopus and Mammalian Cells*

    PubMed Central

    Jevtić, Predrag; Edens, Lisa J.; Li, Xiaoyang; Nguyen, Thang; Chen, Pan; Levy, Daniel L.

    2015-01-01

    A fundamental question in cell biology concerns the regulation of organelle size. While nuclear size is exquisitely controlled in different cell types, inappropriate nuclear enlargement is used to diagnose and stage cancer. Clarifying the functional significance of nuclear size necessitates an understanding of the mechanisms and proteins that control nuclear size. One structural component implicated in the regulation of nuclear morphology is the nuclear lamina, a meshwork of intermediate lamin filaments that lines the inner nuclear membrane. However, there has not been a systematic investigation of how the level and type of lamin expression influences nuclear size, in part due to difficulties in precisely controlling lamin expression levels in vivo. In this study, we circumvent this limitation by studying nuclei in Xenopus laevis egg and embryo extracts, open biochemical systems that allow for precise manipulation of lamin levels by the addition of recombinant proteins. We find that nuclear growth and size are sensitive to the levels of nuclear lamins, with low and high concentrations increasing and decreasing nuclear size, respectively. Interestingly, each type of lamin that we tested (lamins B1, B2, B3, and A) similarly affected nuclear size whether added alone or in combination, suggesting that total lamin concentration, and not lamin type, is more critical to determining nuclear size. Furthermore, we show that altering lamin levels in vivo, both in Xenopus embryos and mammalian tissue culture cells, also impacts nuclear size. These results have implications for normal development and carcinogenesis where both nuclear size and lamin expression levels change. PMID:26429910

  1. Blocking farnesylation of the prelamin A variant in Hutchinson-Gilford progeria syndrome alters the distribution of A-type lamins.

    PubMed

    Wang, Yuexia; Ostlund, Cecilia; Choi, Jason C; Swayne, Theresa C; Gundersen, Gregg G; Worman, Howard J

    2012-01-01

    Mutations in the lamin A/C gene that cause Hutchinson-Gilford progeria syndrome lead to expression of a truncated, permanently farnesylated prelamin A variant called progerin. Blocking farnesylation leads to an improvement in the abnormal nuclear morphology observed in cells expressing progerin, which is associated with a re-localization of the variant protein from the nuclear envelope to the nuclear interior. We now show that a progerin construct that cannot be farnesylated is localized primarily in intranuclear foci and that its diffusional mobility is significantly greater than that of farnesylated progerin localized predominantly at the nuclear envelope. Expression of non-farnesylated progerin in transfected cells leads to a redistribution of lamin A and lamin C away from the nuclear envelope into intranuclear foci but does not significantly affect the localization of endogenous lamin B1 at nuclear envelope. There is a similar redistribution of lamin A and lamin C into intranuclear foci in transfected cells expressing progerin in which protein farnesylation is blocked by treatment with a protein farnesyltransferase inhibitor. Blocking farnesylation of progerin can lead to a redistribution of normal A-type lamins away from the inner nuclear envelope. This may have implications for using drugs that block protein prenylation to treat children with Hutchinson-Gilford progeria syndrome. These findings also provide additional evidence that A-type and B-type lamins can form separate microdomains within the nucleus.

  2. Subcellular localization of SREBP1 depends on its interaction with the C-terminal region of wild-type and disease related A-type lamins

    SciTech Connect

    Duband-Goulet, Isabelle; Woerner, Stephanie; Gasparini, Sylvaine; Attanda, Wikayatou; Konde, Emilie; Tellier-Lebegue, Carine; Craescu, Constantin T.; Roussel, Pascal; Vadrot, Nathalie; Vicart, Patrick; Oestlund, Cecilia; Worman, Howard J.; and others

    2011-12-10

    Lamins A and C are nuclear intermediate filament proteins expressed in most differentiated somatic cells. Previous data suggested that prelamin A, the lamin A precursor, accumulates in some lipodystrophy syndromes caused by mutations in the lamin A/C gene, and binds and inactivates the sterol regulatory element binding protein 1 (SREBP1). Here we show that, in vitro, the tail regions of prelamin A, lamin A and lamin C bind a polypeptide of SREBP1. Such interactions also occur in HeLa cells, since expression of lamin tail regions impedes nucleolar accumulation of the SREBP1 polypeptide fused to a nucleolar localization signal sequence. In addition, the tail regions of A-type lamin variants that occur in Dunnigan-type familial partial lipodystrophy of (R482W) and Hutchison Gilford progeria syndrome ( Increment 607-656) bind to the SREBP1 polypeptide in vitro, and the corresponding FLAG-tagged full-length lamin variants co-immunoprecipitate the SREBP1 polypeptide in cells. Overexpression of wild-type A-type lamins and variants favors SREBP1 polypeptide localization at the intranuclear periphery, suggesting its sequestration. Our data support the hypothesis that variation of A-type lamin protein level and spatial organization, in particular due to disease-linked mutations, influences the sequestration of SREBP1 at the nuclear envelope and thus contributes to the regulation of SREBP1 function.

  3. Subcellular localization of SREBP1 depends on its interaction with the C-terminal region of wild-type and disease related A-type lamins

    PubMed Central

    Duband-Goulet, Isabelle; Woerner, Stephanie; Gasparini, Sylvaine; Attanda, Wikayatou; Kondé, Emilie; Tellier-Lebègue, Carine; Craescu, Constantin T.; Gombault, Aurélie; Roussel, Pascal; Vadrot, Nathalie; Vicart, Patrick; Östlund, Cecilia; Worman, Howard J.; Zinn-Justin, Sophie; Buendia, Brigitte

    2011-01-01

    Lamins A and C are nuclear intermediate filament proteins expressed in most differentiated somatic cells. Previous data suggested that prelamin A, the lamin A precursor, accumulates in some lipodystrophy syndromes caused by mutations in the lamin A/C gene, and binds and inactivates the sterol regulatory element binding protein 1 (SREBP1). Here we show that, in vitro, the tail regions of prelamin A, lamin A and lamin C bind a polypeptide of SREBP1. Such interactions also occur in HeLa cells, since expression of lamin tail regions impedes nucleolar accumulation of the SREBP1 polypeptide fused to a nucleolar localization signal sequence. In addition, the tail regions of A-type lamin variants that occur in Dunnigan-type familial partial lipodystrophy of (R482W) and Hutchison Gilford progeria syndrome (Δ607–656) bind to the SREBP1 polypeptide in vitro, and the corresponding FLAG-tagged full-length lamin variants co-immunoprecipitate the SREBP1 polypeptide in cells. Overexpression of wild-type A-type lamins and variants favors SREBP1 polypeptide localization at the intranuclear periphery, suggesting its sequestration. Our data support the hypothesis that variation of A-type lamin protein level and spatial organization, in particular due to disease-linked mutations, influences the sequestration of SREBP1 at the nuclear envelope and thus contributes to the regulation of SREBP1 function. PMID:21993218

  4. Lamin A-dependent nuclear defects in human aging.

    PubMed

    Scaffidi, Paola; Misteli, Tom

    2006-05-19

    Mutations in the nuclear structural protein lamin A cause the premature aging syndrome Hutchinson-Gilford progeria (HGPS). Whether lamin A plays any role in normal aging is unknown. We show that the same molecular mechanism responsible for HGPS is active in healthy cells. Cell nuclei from old individuals acquire defects similar to those of HGPS patient cells, including changes in histone modifications and increased DNA damage. Age-related nuclear defects are caused by sporadic use, in healthy individuals, of the same cryptic splice site in lamin A whose constitutive activation causes HGPS. Inhibition of this splice site reverses the nuclear defects associated with aging. These observations implicate lamin A in physiological aging.

  5. Concentration-dependent Effects of Nuclear Lamins on Nuclear Size in Xenopus and Mammalian Cells.

    PubMed

    Jevtić, Predrag; Edens, Lisa J; Li, Xiaoyang; Nguyen, Thang; Chen, Pan; Levy, Daniel L

    2015-11-13

    A fundamental question in cell biology concerns the regulation of organelle size. While nuclear size is exquisitely controlled in different cell types, inappropriate nuclear enlargement is used to diagnose and stage cancer. Clarifying the functional significance of nuclear size necessitates an understanding of the mechanisms and proteins that control nuclear size. One structural component implicated in the regulation of nuclear morphology is the nuclear lamina, a meshwork of intermediate lamin filaments that lines the inner nuclear membrane. However, there has not been a systematic investigation of how the level and type of lamin expression influences nuclear size, in part due to difficulties in precisely controlling lamin expression levels in vivo. In this study, we circumvent this limitation by studying nuclei in Xenopus laevis egg and embryo extracts, open biochemical systems that allow for precise manipulation of lamin levels by the addition of recombinant proteins. We find that nuclear growth and size are sensitive to the levels of nuclear lamins, with low and high concentrations increasing and decreasing nuclear size, respectively. Interestingly, each type of lamin that we tested (lamins B1, B2, B3, and A) similarly affected nuclear size whether added alone or in combination, suggesting that total lamin concentration, and not lamin type, is more critical to determining nuclear size. Furthermore, we show that altering lamin levels in vivo, both in Xenopus embryos and mammalian tissue culture cells, also impacts nuclear size. These results have implications for normal development and carcinogenesis where both nuclear size and lamin expression levels change. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins

    NASA Technical Reports Server (NTRS)

    Schatten, G.; Schatten, H.; Simerly, C.; Maul, G. G.; Chaly, N.

    1985-01-01

    Nuclear structural changes during fertilization and embryogenesis in mice and sea urchins are traced using four antibodies. The oocytes from virgin female mice, morulae and blastocytes from mated females, and gametes from the sea urchin Lytechnius variegatis are studied using mouse monoclonal antibodies to nuclear lamin A/C, monoclonal antibody to P1, human autoimmune antibodies to lamin A/C, and to lamin B. The mouse fertilization data reveal no lamins on the oocyte; however, lamins are present on the pronuclei, and chromosomes are found on the oocytes and pronuclei. It is detected that on the sea urchin sperm the lamins are reduced to acrosomal and centriolar fossae and peripheral antigens are around the sperm nucleus. The mouse sperm bind lamin antibodies regionally and do not contain antigens. Lamins and antigens are observed on both pronuclei and chromosomes during sea urchin fertilization. Mouse embryogenesis reveals that lamin A/C is not recognized at morula and blastocyst stages; however, lamin B stains are retained. In sea urchin embryogenesis lamin recognition is lost at the blastrula, gastrula, and plutei stages. It is noted that nuclear lamins lost during spermatogenesis are restored at fertilization and peripheral antigens are associated with the surface of chromosomes during meiosis and mitosis and with the periphery of the pronuclei and nuclei during interphase.

  7. Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins

    NASA Technical Reports Server (NTRS)

    Schatten, G.; Schatten, H.; Simerly, C.; Maul, G. G.; Chaly, N.

    1985-01-01

    Nuclear structural changes during fertilization and embryogenesis in mice and sea urchins are traced using four antibodies. The oocytes from virgin female mice, morulae and blastocytes from mated females, and gametes from the sea urchin Lytechnius variegatis are studied using mouse monoclonal antibodies to nuclear lamin A/C, monoclonal antibody to P1, human autoimmune antibodies to lamin A/C, and to lamin B. The mouse fertilization data reveal no lamins on the oocyte; however, lamins are present on the pronuclei, and chromosomes are found on the oocytes and pronuclei. It is detected that on the sea urchin sperm the lamins are reduced to acrosomal and centriolar fossae and peripheral antigens are around the sperm nucleus. The mouse sperm bind lamin antibodies regionally and do not contain antigens. Lamins and antigens are observed on both pronuclei and chromosomes during sea urchin fertilization. Mouse embryogenesis reveals that lamin A/C is not recognized at morula and blastocyst stages; however, lamin B stains are retained. In sea urchin embryogenesis lamin recognition is lost at the blastrula, gastrula, and plutei stages. It is noted that nuclear lamins lost during spermatogenesis are restored at fertilization and peripheral antigens are associated with the surface of chromosomes during meiosis and mitosis and with the periphery of the pronuclei and nuclei during interphase.

  8. Nuclear lamins and peripheral nuclear antigens during fertilization and embryogenesis in mice and sea urchins

    SciTech Connect

    Schatten, G.; Schatten, H.; Simerly, C.; Maul, G.G.; Chaly, N.

    1985-07-01

    Nuclear structural changes during fertilization and embryogenesis in mice and sea urchins are traced using four antibodies. The oocytes from virgin female mice, morulae and blastocytes from mated females, and gametes from the sea urchin Lytechnius variegatis are studied using mouse monoclonal antibodies to nuclear lamin A/C, monoclonal antibody to P1, human autoimmune antibodies to lamin A/C, and to lamin B. The mouse fertilization data reveal no lamins on the oocyte; however, lamins are present on the pronuclei, and chromosomes are found on the oocytes and pronuclei. It is detected that on the sea urchin sperm the lamins are reduced to acrosomal and centriolar fossae and peripheral antigens are around the sperm nucleus. The mouse sperm bind lamin antibodies regionally and do not contain antigens. Lamins and antigens are observed on both pronuclei and chromosomes during sea urchin fertilization. Mouse embryogenesis reveals that lamin A/C is not recognized at morula and blastocyst stages; however, lamin B stains are retained. In sea urchin embryogenesis lamin recognition is lost at the blastrula, gastrula, and plutei stages. It is noted that nuclear lamins lost during spermatogenesis are restored at fertilization and peripheral antigens are associated with the surface of chromosomes during meiosis and mitosis and with the periphery of the pronuclei and nuclei during interphase. 32 references.

  9. Nuclear lamins during gametogenesis, fertilization and early development

    NASA Technical Reports Server (NTRS)

    Maul, G. G.; Schatten, G.

    1986-01-01

    The distribution of lamins (described by Gerace, 1978, as major proteins of nuclear envelope) during gametogenesis, fertilization, and early development was investigated in germ cells of a mouse (Mus musculus), an echinoderm (Lytechinus variegatus), and the surf clam (Spisula solidissima) was investigated in order to determine whether the differences detected could be correlated with differences in the function of cells in these stages of the germ cells. In order to monitor the behavior of lamins, the gametes and embryos were labeled with antibodies to lamins A, C, and B extracted from autoimmune sera of patients with scleroderma and Lupus erythematosus. Results indicated that lamin B could be identified in nuclear envelopes on only those nuclei where chromatin is attached and where RNA synthesis takes place.

  10. Nuclear lamins during gametogenesis, fertilization and early development

    NASA Technical Reports Server (NTRS)

    Maul, G. G.; Schatten, G.

    1986-01-01

    The distribution of lamins (described by Gerace, 1978, as major proteins of nuclear envelope) during gametogenesis, fertilization, and early development was investigated in germ cells of a mouse (Mus musculus), an echinoderm (Lytechinus variegatus), and the surf clam (Spisula solidissima) was investigated in order to determine whether the differences detected could be correlated with differences in the function of cells in these stages of the germ cells. In order to monitor the behavior of lamins, the gametes and embryos were labeled with antibodies to lamins A, C, and B extracted from autoimmune sera of patients with scleroderma and Lupus erythematosus. Results indicated that lamin B could be identified in nuclear envelopes on only those nuclei where chromatin is attached and where RNA synthesis takes place.

  11. Fate of the inner nuclear membrane protein lamin B receptor and nuclear lamins in herpes simplex virus type 1 infection.

    PubMed

    Scott, E S; O'Hare, P

    2001-09-01

    During herpesvirus egress, capsids bud through the inner nuclear membrane. Underlying this membrane is the nuclear lamina, a meshwork of intermediate filaments with which it is tightly associated. Details of alterations to the lamina and the inner nuclear membrane during infection and the mechanisms involved in capsid transport across these structures remain unclear. Here we describe the fate of key protein components of the nuclear envelope and lamina during herpes simplex virus type 1 (HSV-1) infection. We followed the distribution of the inner nuclear membrane protein lamin B receptor (LBR) and lamins A and B(2) tagged with green fluorescent protein (GFP) in live infected cells. Together with additional results from indirect immunofluorescence, our studies reveal major morphologic distortion of nuclear-rim LBR and lamins A/C, B(1), and B(2). By 8 h p.i., we also observed a significant redistribution of LBR-GFP to the endoplasmic reticulum, where it colocalized with a subpopulation of cytoplasmic glycoprotein B by immunofluorescence. In addition, analysis by fluorescence recovery after photobleaching reveals that LBR-GFP exhibited increased diffusional mobility within the nuclear membrane of infected cells. This is consistent with the disruption of interactions between LBR and the underlying lamina. In addition to studying stably expressed GFP-lamins by fluorescence microscopy, we studied endogenous A- and B-type lamins in infected cells by Western blotting. Both approaches reveal a loss of lamins associated with virus infection. These data indicate major disruption of the nuclear envelope and lamina of HSV-1-infected cells and are consistent with a virus-induced dismantling of the nuclear lamina, possibly in order to gain access to the inner nuclear membrane.

  12. Post-natal myogenic and adipogenic developmental: defects and metabolic impairment upon loss of A-type lamins.

    PubMed

    Kubben, Nard; Voncken, Jan Willem; Konings, Gonda; van Weeghel, Michel; van den Hoogenhof, Maarten Mg; Gijbels, Marion; van Erk, Arie; Schoonderwoerd, Kees; van den Bosch, Bianca; Dahlmans, Vivian; Calis, Chantal; Houten, Sander M; Misteli, Tom; Pinto, Yigal M

    2011-01-01

    A-type lamins are a major component of the nuclear lamina. Mutations in the LMNA gene, which encodes the A-type lamins A and C, cause a set of phenotypically diverse diseases collectively called laminopathies. While adult LMNA null mice show various symptoms typically associated with laminopathies, the effect of loss of lamin A/C on early post-natal development is poorly understood. Here we developed a novel LMNA null mouse (LMNA(GT-/-)) based on genetrap technology and analyzed its early post-natal development. We detect LMNA transcripts in heart, the outflow tract, dorsal aorta, liver and somites during early embryonic development. Loss of A-type lamins results in severe growth retardation and developmental defects of the heart, including impaired myocyte hypertrophy, skeletal muscle hypotrophy, decreased amounts of subcutaneous adipose tissue and impaired ex vivo adipogenic differentiation. These defects cause death at 2 to 3 weeks post partum associated with muscle weakness and metabolic complications, but without the occurrence of dilated cardiomyopathy or an obvious progeroid phenotype. Our results indicate that defective early post-natal development critically contributes to the disease phenotypes in adult laminopathies.

  13. Roles of endothelial A-type lamins in migration of T cells on and under endothelial layers

    NASA Astrophysics Data System (ADS)

    Song, Kwang Hoon; Lee, Jaehyun; Park, Hyoungjun; Kim, Hye Mi; Park, Jeehun; Kwon, Keon Woo; Doh, Junsang

    2016-03-01

    Stiff nuclei in cell-dense microenvironments may serve as distinct biomechanical cues for cell migration, but such a possibility has not been tested experimentally. As a first step addressing this question, we altered nuclear stiffness of endothelial cells (ECs) by reducing the expression of A-type lamins using siRNA, and investigated the migration of T cells on and under EC layers. While most T cells crawling on control EC layers avoided crossing over EC nuclei, a significantly higher fraction of T cells on EC layers with reduced expression of A-type lamins crossed over EC nuclei. This result suggests that stiff EC nuclei underlying T cells may serve as “duro-repulsive” cues to direct T cell migration toward less stiff EC cytoplasm. During subendothelial migration under EC layers with reduced expression of A-type lamins, T cells made prolonged contact and substantially deformed EC nuclei, resulting in reduced speed and directional persistence. This result suggests that EC nuclear stiffness promotes fast and directionally persistent subendothelial migration of T cells by allowing minimum interaction between T cells and EC nuclei.

  14. Roles of endothelial A-type lamins in migration of T cells on and under endothelial layers

    PubMed Central

    Song, Kwang Hoon; Lee, Jaehyun; Park, HyoungJun; Kim, Hye Mi; Park, Jeehun; Kwon, Keon Woo; Doh, Junsang

    2016-01-01

    Stiff nuclei in cell-dense microenvironments may serve as distinct biomechanical cues for cell migration, but such a possibility has not been tested experimentally. As a first step addressing this question, we altered nuclear stiffness of endothelial cells (ECs) by reducing the expression of A-type lamins using siRNA, and investigated the migration of T cells on and under EC layers. While most T cells crawling on control EC layers avoided crossing over EC nuclei, a significantly higher fraction of T cells on EC layers with reduced expression of A-type lamins crossed over EC nuclei. This result suggests that stiff EC nuclei underlying T cells may serve as “duro-repulsive” cues to direct T cell migration toward less stiff EC cytoplasm. During subendothelial migration under EC layers with reduced expression of A-type lamins, T cells made prolonged contact and substantially deformed EC nuclei, resulting in reduced speed and directional persistence. This result suggests that EC nuclear stiffness promotes fast and directionally persistent subendothelial migration of T cells by allowing minimum interaction between T cells and EC nuclei. PMID:26996137

  15. Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin

    PubMed Central

    Dechat, Thomas; Pfleghaar, Katrin; Sengupta, Kaushik; Shimi, Takeshi; Shumaker, Dale K.; Solimando, Liliana; Goldman, Robert D.

    2008-01-01

    Over the past few years it has become evident that the intermediate filament proteins, the types A and B nuclear lamins, not only provide a structural framework for the nucleus, but are also essential for many aspects of normal nuclear function. Insights into lamin-related functions have been derived from studies of the remarkably large number of disease-causing mutations in the human lamin A gene. This review provides an up-to-date overview of the functions of nuclear lamins, emphasizing their roles in epigenetics, chromatin organization, DNA replication, transcription, and DNA repair. In addition, we discuss recent evidence supporting the importance of lamins in viral infections. PMID:18381888

  16. Prelamin A and lamin A appear to be dispensable in the nuclear lamina.

    PubMed

    Fong, Loren G; Ng, Jennifer K; Lammerding, Jan; Vickers, Timothy A; Meta, Margarita; Coté, Nathan; Gavino, Bryant; Qiao, Xin; Chang, Sandy Y; Young, Stephanie R; Yang, Shao H; Stewart, Colin L; Lee, Richard T; Bennett, C Frank; Bergo, Martin O; Young, Stephen G

    2006-03-01

    Lamin A and lamin C, both products of Lmna, are key components of the nuclear lamina. In the mouse, a deficiency in both lamin A and lamin C leads to slow growth, muscle weakness, and death by 6 weeks of age. Fibroblasts deficient in lamins A and C contain misshapen and structurally weakened nuclei, and emerin is mislocalized away from the nuclear envelope. The physiologic rationale for the existence of the 2 different Lmna products lamin A and lamin C is unclear, although several reports have suggested that lamin A may have particularly important functions, for example in the targeting of emerin and lamin C to the nuclear envelope. Here we report the development of lamin C-only mice (Lmna(LCO/LCO)), which produce lamin C but no lamin A or prelamin A (the precursor to lamin A). Lmna(LCO/LCO) mice were entirely healthy, and Lmna(LCO/LCO) cells displayed normal emerin targeting and exhibited only very minimal alterations in nuclear shape and nuclear deformability. Thus, at least in the mouse, prelamin A and lamin A appear to be dispensable. Nevertheless, an accumulation of farnesyl-prelamin A (as occurs with a deficiency in the prelamin A processing enzyme Zmpste24) caused dramatically misshapen nuclei and progeria-like disease phenotypes. The apparent dispensability of prelamin A suggested that lamin A-related progeroid syndromes might be treated with impunity by reducing prelamin A synthesis. Remarkably, the presence of a single Lmna(LCO) allele eliminated the nuclear shape abnormalities and progeria-like disease phenotypes in Zmpste24-/- mice. Moreover, treating Zmpste24-/- cells with a prelamin A-specific antisense oligonucleotide reduced prelamin A levels and significantly reduced the frequency of misshapen nuclei. These studies suggest a new therapeutic strategy for treating progeria and other lamin A diseases.

  17. Prelamin A and lamin A appear to be dispensable in the nuclear lamina

    PubMed Central

    Fong, Loren G.; Ng, Jennifer K.; Lammerding, Jan; Vickers, Timothy A.; Meta, Margarita; Coté, Nathan; Gavino, Bryant; Qiao, Xin; Chang, Sandy Y.; Young, Stephanie R.; Yang, Shao H.; Stewart, Colin L.; Lee, Richard T.; Bennett, C. Frank; Bergo, Martin O.; Young, Stephen G.

    2006-01-01

    Lamin A and lamin C, both products of Lmna, are key components of the nuclear lamina. In the mouse, a deficiency in both lamin A and lamin C leads to slow growth, muscle weakness, and death by 6 weeks of age. Fibroblasts deficient in lamins A and C contain misshapen and structurally weakened nuclei, and emerin is mislocalized away from the nuclear envelope. The physiologic rationale for the existence of the 2 different Lmna products lamin A and lamin C is unclear, although several reports have suggested that lamin A may have particularly important functions, for example in the targeting of emerin and lamin C to the nuclear envelope. Here we report the development of lamin C–only mice (Lmna+/+), which produce lamin C but no lamin A or prelamin A (the precursor to lamin A). Lmna+/+ mice were entirely healthy, and Lmna+/+ cells displayed normal emerin targeting and exhibited only very minimal alterations in nuclear shape and nuclear deformability. Thus, at least in the mouse, prelamin A and lamin A appear to be dispensable. Nevertheless, an accumulation of farnesyl–prelamin A (as occurs with a deficiency in the prelamin A processing enzyme Zmpste24) caused dramatically misshapen nuclei and progeria-like disease phenotypes. The apparent dispensability of prelamin A suggested that lamin A–related progeroid syndromes might be treated with impunity by reducing prelamin A synthesis. Remarkably, the presence of a single LmnaLCO allele eliminated the nuclear shape abnormalities and progeria-like disease phenotypes in Zmpste24–/– mice. Moreover, treating Zmpste24–/– cells with a prelamin A–specific antisense oligonucleotide reduced prelamin A levels and significantly reduced the frequency of misshapen nuclei. These studies suggest a new therapeutic strategy for treating progeria and other lamin A diseases. PMID:16511604

  18. A-type lamins bind both hetero- and euchromatin, the latter being regulated by lamina-associated polypeptide 2 alpha

    PubMed Central

    Gesson, Kevin; Rescheneder, Philipp; Skoruppa, Michael P.; von Haeseler, Arndt; Dechat, Thomas

    2016-01-01

    Lamins are components of the peripheral nuclear lamina and interact with heterochromatic genomic regions, termed lamina-associated domains (LADs). In contrast to lamin B1 being primarily present at the nuclear periphery, lamin A/C also localizes throughout the nucleus, where it associates with the chromatin-binding protein lamina-associated polypeptide (LAP) 2 alpha. Here, we show that lamin A/C also interacts with euchromatin, as determined by chromatin immunoprecipitation of euchromatin- and heterochromatin-enriched samples. By way of contrast, lamin B1 was only found associated with heterochromatin. Euchromatic regions occupied by lamin A/C overlap with those bound by LAP2alpha, and lack of LAP2alpha in LAP2alpha-deficient cells shifts binding of lamin A/C toward more heterochromatic regions. These alterations in lamin A/C-chromatin interactions correlate with changes in epigenetic histone marks in euchromatin but do not significantly affect gene expression. Loss of lamin A/C in heterochromatic regions in LAP2alpha-deficient cells, however, correlated with increased gene expression. Our data show a novel role of nucleoplasmic lamin A/C and LAP2alpha in regulating euchromatin. PMID:26798136

  19. Dynamics of lamin A/C in porcine embryos produced by nuclear transfer.

    PubMed

    Lee, Kiho; Fodor, William L; Machaty, Zoltan

    2007-09-01

    This study was conducted to investigate the presence of lamin A/C in porcine nuclear transfer embryos and to determine whether lamin A/C can serve as a potential marker for nuclear reprogramming. First, lamin A/C was studied in oocytes and embryos produced by fertilization or parthenogenetic oocyte activation. We found that lamin A/C was present in the nuclear lamina of oocytes at the germinal vesicle stage while it was absent in mature oocytes. Lamin A/C was detected throughout preimplantation development in both in vivo-derived and parthenogenetic embryos. Incubation of the activated oocytes in the presence of alpha-amanitin (an inhibitor of RNA polymerase II), or cycloheximide (a protein synthesis inhibitor) did not perturb lamin A/C assembly, indicating that the assembly resulted from solubilized lamins dispersed in the cytoplasm. In nuclear transfer embryos, the lamin A/C signal that had previously been identified in fibroblast nuclei disappeared soon after fusion. It became detectable again after the formation of the pronucleus-like structure, and all nuclear transfer embryos displayed lamin A/C staining during early development. Olfactory bulb progenitor cells lacked lamin A/C; however, when such cells were fused with enucleated oocytes, the newly formed nuclear envelopes stained positive for lamin A/C. These findings suggest that recipient oocytes remodel the donor nuclei using type A lamins dispersed in the ooplasm. The results also indicate that lamin A/C is present in the nuclear envelope of pig oocytes and early embryos and unlike in some other species, its presence after nuclear transfer is not an indicator of erroneous reprogramming.

  20. Laminates

    NASA Astrophysics Data System (ADS)

    Lepedat, Karin; Wagner, Robert; Lang, Jürgen

    The use of phenolic resin for the impregnation of a carrier material such as paper or fabric based on either organic or inorganic fibers was and still is one of the most important application areas for liquid phenolic resins. Substrates like paper, cotton, or glass fabric impregnated with phenolic resins are used as core layers for decorative and technical laminates and for many other different industrial applications. Nowadays, phenolic resins for decorative laminates used for furniture, flooring, or in the construction and transportation industry have gained significant market share. The Laminates chapter mainly describes the manufacture of decorative laminates especially the impregnation and pressing process with special emphasis to new technological developments and recent trends. Moreover, the different types of laminates are introduced, combined with some brief comments as they relate to the market for decorative surfaces.

  1. Differential basal-to-apical accessibility of lamin A/C epitopes in the nuclear lamina regulated by changes in cytoskeletal tension

    NASA Astrophysics Data System (ADS)

    Ihalainen, Teemu O.; Aires, Lina; Herzog, Florian A.; Schwartlander, Ruth; Moeller, Jens; Vogel, Viola

    2015-12-01

    Nuclear lamins play central roles at the intersection between cytoplasmic signalling and nuclear events. Here, we show that at least two N- and C-terminal lamin epitopes are not accessible at the basal side of the nuclear envelope under environmental conditions known to upregulate cell contractility. The conformational epitope on the Ig-domain of A-type lamins is more buried in the basal than apical nuclear envelope of human mesenchymal stem cells undergoing osteogenesis (but not adipogenesis), and in fibroblasts adhering to rigid (but not soft) polyacrylamide hydrogels. This structural polarization of the lamina is promoted by compressive forces, emerges during cell spreading, and requires lamin A/C multimerization, intact nucleoskeleton-cytoskeleton linkages (LINC), and apical-actin stress-fibre assembly. Notably, the identified Ig-epitope overlaps with emerin, DNA and histone binding sites, and comprises various laminopathy mutation sites. Our findings should help decipher how the physical properties of cellular microenvironments regulate nuclear events.

  2. Differential basal-to-apical accessibility of lamin A/C epitopes in the nuclear lamina regulated by changes in cytoskeletal tension

    PubMed Central

    Herzog, Florian A.; Schwartlander, Ruth; Moeller, Jens; Vogel, Viola

    2015-01-01

    Nuclear lamins play central roles at the intersection between cytoplasmic signalling and nuclear events. Here, we show that at least two N- and C-terminal lamin epitopes are not accessible at the basal side of the nuclear envelope under environmental conditions known to upregulate cell contractility. The conformational epitope on the Ig-domain of A-type lamins is more buried in the basal than apical nuclear envelope of human mesenchymal stem cells undergoing osteogenesis (but not adipogenesis), and in fibroblasts adhering to rigid (but not soft) polyacrylamide hydrogels. This structural polarization of the lamina is promoted by compressive forces, emerges during cell spreading, and requires lamin A/C multimerization, intact nucleoskeleton-cytoskeleton linkages (LINC), and apical-actin stress-fibre assembly. Notably, the identified Igepitope overlaps with emerin, DNA and histone binding sites, and comprises various laminopathy mutation sites. Our findings should help deciphering how the physical properties of cellular microenvironments regulate nuclear events. PMID:26301768

  3. Lamin A and microtubules collaborate to maintain nuclear morphology.

    PubMed

    Tariq, Zeshan; Zhang, Haoyue; Chia-Liu, Alexander; Shen, Yang; Gete, Yantenew; Xiong, Zheng-Mei; Tocheny, Claire; Campanello, Leonard; Wu, Di; Losert, Wolfgang; Cao, Kan

    2017-07-04

    Lamin A (LA) is a critical structural component of the nuclear lamina. Mutations within the LA gene (LMNA) lead to several human disorders, most striking of which is Hutchinson-Gilford Progeria Syndrome (HGPS), a premature aging disorder. HGPS cells are best characterized by an abnormal nuclear morphology known as nuclear blebbing, which arises due to the accumulation of progerin, a dominant mutant form of LA. The microtubule (MT) network is known to mediate changes in nuclear morphology in the context of specific events such as mitosis, cell polarization, nucleus positioning and cellular migration. What is less understood is the role of the microtubule network in determining nuclear morphology during interphase. In this study, we elucidate the role of the cytoskeleton in regulation and misregulation of nuclear morphology through perturbations of both the lamina and the microtubule network. We found that LA knockout cells exhibit a crescent shape morphology associated with the microtubule-organizing center. Furthermore, this crescent shape ameliorates upon treatment with MT drugs, Nocodazole or Taxol. Expression of progerin, in LA knockout cells also rescues the crescent shape, although the response to Nocodazole or Taxol treatment is altered in comparison to cells expressing LA. Together these results describe a collaborative effort between LA and the MT network to maintain nuclear morphology.

  4. Laminopathic mutations interfere with the assembly, localization, and dynamics of nuclear lamins.

    PubMed

    Wiesel, Naama; Mattout, Anna; Melcer, Shai; Melamed-Book, Naomi; Herrmann, Harald; Medalia, Ohad; Aebi, Ueli; Gruenbaum, Yosef

    2008-01-08

    Lamins are nuclear intermediate filament proteins and the major building blocks of the nuclear lamina. Besides providing nuclear shape and mechanical stability, lamins are required for chromatin organization, transcription regulation, DNA replication, nuclear assembly, nuclear positioning, and apoptosis. Mutations in human lamins cause many different heritable diseases, affecting various tissues and causing early aging. Although many of these mutations result in nuclear deformation, their effects on lamin filament assembly are unknown. Caenorhabditis elegans has a single evolutionarily conserved lamin protein, which can form stable 10-nm-thick filaments in vitro. To gain insight into the molecular basis of lamin filament assembly and the effects of laminopathic mutations on this process, we investigated mutations in conserved residues of the rod and tail domains that are known to cause various laminopathies in human. We show that 8 of 14 mutant lamins present WT-like assembly into filaments or paracrystals, whereas 6 mutants show assembly defects. Correspondingly, expressing these mutants in transgenic animals shows abnormal distribution of Ce-lamin, abnormal nuclear shape or change in lamin mobility. These findings help in understanding the role of individual residues and domains in laminopathy pathology and, eventually, promote the development of therapeutic interventions.

  5. Gene-rich chromosomal regions are preferentially localized in the lamin B deficient nuclear blebs of atypical progeria cells.

    PubMed

    Bercht Pfleghaar, Katrin; Taimen, Pekka; Butin-Israeli, Veronika; Shimi, Takeshi; Langer-Freitag, Sabine; Markaki, Yolanda; Goldman, Anne E; Wehnert, Manfred; Goldman, Robert D

    2015-01-01

    More than 20 mutations in the gene encoding A-type lamins (LMNA) cause progeria, a rare premature aging disorder. The major pathognomonic hallmarks of progeria cells are seen as nuclear deformations or blebs that are related to the redistribution of A- and B-type lamins within the nuclear lamina. However, the functional significance of these progeria-associated blebs remains unknown. We have carried out an analysis of the structural and functional consequences of progeria-associated nuclear blebs in dermal fibroblasts from a progeria patient carrying a rare point mutation p.S143F (C428T) in lamin A/C. These blebs form microdomains that are devoid of major structural components of the nuclear envelope (NE)/lamina including B-type lamins and nuclear pore complexes (NPCs) and are enriched in A-type lamins. Using laser capture microdissection and comparative genomic hybridization (CGH) analyses, we show that, while these domains are devoid of centromeric heterochromatin and gene-poor regions of chromosomes, they are enriched in gene-rich chromosomal regions. The active form of RNA polymerase II is also greatly enriched in blebs as well as nascent RNA but the nuclear co-activator SKIP is significantly reduced in blebs compared to other transcription factors. Our results suggest that the p.S143F progeria mutation has a severe impact not only on the structure of the lamina but also on the organization of interphase chromatin domains and transcription. These structural defects are likely to contribute to gene expression changes reported in progeria and other types of laminopathies.

  6. Gene-rich chromosomal regions are preferentially localized in the lamin B deficient nuclear blebs of atypical progeria cells

    PubMed Central

    Bercht Pfleghaar, Katrin; Taimen, Pekka; Butin-Israeli, Veronika; Shimi, Takeshi; Langer-Freitag, Sabine; Markaki, Yolanda; Goldman, Anne E; Wehnert, Manfred; Goldman, Robert D

    2015-01-01

    More than 20 mutations in the gene encoding A-type lamins (LMNA) cause progeria, a rare premature aging disorder. The major pathognomonic hallmarks of progeria cells are seen as nuclear deformations or blebs that are related to the redistribution of A- and B-type lamins within the nuclear lamina. However, the functional significance of these progeria-associated blebs remains unknown. We have carried out an analysis of the structural and functional consequences of progeria-associated nuclear blebs in dermal fibroblasts from a progeria patient carrying a rare point mutation p.S143F (C428T) in lamin A/C. These blebs form microdomains that are devoid of major structural components of the nuclear envelope (NE)/lamina including B-type lamins and nuclear pore complexes (NPCs) and are enriched in A-type lamins. Using laser capture microdissection and comparative genomic hybridization (CGH) analyses, we show that, while these domains are devoid of centromeric heterochromatin and gene-poor regions of chromosomes, they are enriched in gene-rich chromosomal regions. The active form of RNA polymerase II is also greatly enriched in blebs as well as nascent RNA but the nuclear co-activator SKIP is significantly reduced in blebs compared to other transcription factors. Our results suggest that the p.S143F progeria mutation has a severe impact not only on the structure of the lamina but also on the organization of interphase chromatin domains and transcription. These structural defects are likely to contribute to gene expression changes reported in progeria and other types of laminopathies. PMID:25738644

  7. A-type lamins and cardiovascular disease in premature aging syndromes.

    PubMed

    Dorado, Beatriz; Andrés, Vicente

    2017-01-10

    Lamin A is a nuclear intermediate filament protein with important structural and regulatory roles in most differentiated mammalian cells. Excessive accumulation of its precursor prelamin A or the mutant form called 'progerin' causes premature aging syndromes. Progeroid 'laminopathies' are characterized by severe cardiovascular problems (cardiac electrical defects, vascular calcification and stiffening, atherosclerosis, myocardial infarction, and stroke) and premature death. Here, we review studies in cell and mouse models and patients that are unraveling how abnormal prelamin A and progerin accumulation accelerates cardiovascular disease and aging. This knowledge is essential for developing effective therapies to treat progeria and may help identify new mechanisms underlying normal aging.

  8. Sorting Nexin 6 Enhances Lamin A Synthesis and Incorporation into the Nuclear Envelope

    PubMed Central

    González-Granado, Jose M.; Navarro-Puche, Ana; Molina-Sanchez, Pedro; Blanco-Berrocal, Marta; Viana, Rosa; de Mora, Jaime Font; Andrés, Vicente

    2014-01-01

    Nuclear lamins are important structural and functional proteins in mammalian cells, but little is known about the mechanisms and cofactors that regulate their traffic into the nucleus. Here, we demonstrate that trafficking of lamin A, but not lamin B1, and its assembly into the nuclear envelope are regulated by sorting nexin 6 (SNX6), a major component of the retromer that targets proteins and other molecules to specific subcellular locations. SNX6 interacts with lamin A in vitro and in vivo and links it to the outer surface of the endoplasmic reticulum in human and mouse cells. SNX6 transports its lamin A cargo to the nuclear envelope in a process that takes several hours. Lamin A protein levels in the nucleus augment or decrease, respectively, upon gain or loss of SNX6 function. We further show that SNX6-dependent lamin A nuclear import occurs across the nuclear pore complex via a RAN-GTP-dependent mechanism. These results identify SNX6 as a key regulator of lamin A synthesis and incorporation into the nuclear envelope. PMID:25535984

  9. Nuclear lamin-A scales with tissue stiffness and enhances matrix-directed differentiation.

    PubMed

    Swift, Joe; Ivanovska, Irena L; Buxboim, Amnon; Harada, Takamasa; Dingal, P C Dave P; Pinter, Joel; Pajerowski, J David; Spinler, Kyle R; Shin, Jae-Won; Tewari, Manorama; Rehfeldt, Florian; Speicher, David W; Discher, Dennis E

    2013-08-30

    Tissues can be soft like fat, which bears little stress, or stiff like bone, which sustains high stress, but whether there is a systematic relationship between tissue mechanics and differentiation is unknown. Here, proteomics analyses revealed that levels of the nucleoskeletal protein lamin-A scaled with tissue elasticity, E, as did levels of collagens in the extracellular matrix that determine E. Stem cell differentiation into fat on soft matrix was enhanced by low lamin-A levels, whereas differentiation into bone on stiff matrix was enhanced by high lamin-A levels. Matrix stiffness directly influenced lamin-A protein levels, and, although lamin-A transcription was regulated by the vitamin A/retinoic acid (RA) pathway with broad roles in development, nuclear entry of RA receptors was modulated by lamin-A protein. Tissue stiffness and stress thus increase lamin-A levels, which stabilize the nucleus while also contributing to lineage determination.

  10. Nuclear Lamin-A Scales with Tissue Stiffness and Enhances Matrix-Directed Differentiation

    PubMed Central

    Swift, Joe; Ivanovska, Irena L.; Buxboim, Amnon; Harada, Takamasa; Dingal, P. C. Dave P.; Pinter, Joel; Pajerowski, J. David; Spinler, Kyle R.; Shin, Jae-Won; Tewari, Manorama; Rehfeldt, Florian; Speicher, David W.; Discher, Dennis E.

    2014-01-01

    Tissues can be soft like fat, which bears little stress, or stiff like bone, which sustains high stress, but whether there is a systematic relationship between tissue mechanics and differentiation is unknown. Here, proteomics analyses revealed that levels of the nucleoskeletal protein lamin-A scaled with tissue elasticity, E, as did levels of collagens in the extracellular matrix that determine E. Stem cell differentiation into fat on soft matrix was enhanced by low lamin-A levels, whereas differentiation into bone on stiff matrix was enhanced by high lamin-A levels. Matrix stiffness directly influenced lamin-A protein levels, and, although lamin-A transcription was regulated by the vitamin A/retinoic acid (RA) pathway with broad roles in development, nuclear entry of RA receptors was modulated by lamin-A protein. Tissue stiffness and stress thus increase lamin-A levels, which stabilize the nucleus while also contributing to lineage determination. PMID:23990565

  11. The nuclear membrane and mechanotransduction: impaired nuclear mechanics and mechanotransduction in lamin A/C deficient cells.

    PubMed

    Lammerding, Jan; Lee, Richard T

    2005-01-01

    Mutations in the lamin A/C gene cause a variety of human diseases including Emery-Dreifuss muscular dystrophy, dilated cardiomyopathy and Hutchinson-Gilford progeria syndrome. The tissue specific effects of lamin mutations are unclear, in part because the function of lamin A/C is incompletely defined, but the many muscle specific phenotypes suggest that defective lamin A/C could increase cellular mechanical sensitivity. Lamin A/C deficient fibroblasts were subjected to mechanical strain to measure nuclear mechanical properties and strain-induced signalling. We found that lamin A/C deficient fibroblasts are characterized by impaired nuclear mechanics and mechanotransduction, reflected by increased nuclear deformations, increased nuclear fragility, attenuated expression of mechanosensitive genes, and impaired transcriptional activation, leading to impaired viability of mechanically strained cells. Lamins and other nuclear envelope proteins can thus affect several levels of the mechanotransduction cascade, altering nuclear and cytoskeletal mechanics as well as playing an important role in mechanically activated gene regulation. Individual mutations in the lamin A/C gene could potentially selectively interfere with any of these functions, explaining the tissue-specific effects observed in the laminopathies.

  12. Nuclear envelope localization of LEMD2 is developmentally dynamic and lamin A/C dependent yet insufficient for heterochromatin tethering.

    PubMed

    Thanisch, Katharina; Song, Congdi; Engelkamp, Dieter; Koch, Jeannette; Wang, Audrey; Hallberg, Einar; Foisner, Roland; Leonhardt, Heinrich; Stewart, Colin L; Joffe, Boris; Solovei, Irina

    Peripheral heterochromatin in mammalian nuclei is tethered to the nuclear envelope by at least two mechanisms here referred to as the A- and B-tethers. The A-tether includes lamins A/C and additional unknown components presumably INM protein(s) interacting with both lamins A/C and chromatin. The B-tether includes the inner nuclear membrane (INM) protein Lamin B-receptor, which binds B-type lamins and chromatin. Generally, at least one of the tethers is always present in the nuclear envelope of mammalian cells. Deletion of both causes the loss of peripheral heterochromatin and consequently inversion of the entire nuclear architecture, with this occurring naturally in rod photoreceptors of nocturnal mammals. The tethers are differentially utilized during development, regulate gene expression in opposite manners, and play an important role during cell differentiation. Here we aimed to identify the unknown chromatin binding component(s) of the A-tether. We analyzed 10 mouse tissues by immunostaining with antibodies against 7 INM proteins and found that every cell type has specific, although differentially and developmentally regulated, sets of these proteins. In particular, we found that INM protein LEMD2 is concomitantly expressed with A-type lamins in various cell types but is lacking in inverted nuclei of rod cells. Truncation or deletion of Lmna resulted in the downregulation and mislocalization of LEMD2, suggesting that the two proteins interact and pointing at LEMD2 as a potential chromatin binding mediator of the A-tether. Using nuclei of mouse rods as an experimental model lacking peripheral heterochromatin, we expressed a LEMD2 transgene alone or in combination with lamin C in these cells and observed no restoration of peripheral heterochromatin in either case. We conclude that in contrary to the B-tether, the A-tether has a more intricate composition and consists of multiple components that presumably vary, at differing degrees of redundancy, between cell

  13. Herpes simplex virus 2 UL13 protein kinase disrupts nuclear lamins

    SciTech Connect

    Cano-Monreal, Gina L.; Wylie, Kristine M.; Cao, Feng; Tavis, John E.; Morrison, Lynda A.

    2009-09-15

    Herpesviruses must cross the inner nuclear membrane and underlying lamina to exit the nucleus. HSV-1 US3 and PKC can phosphorylate lamins and induce their dispersion but do not elicit all of the phosphorylated lamin species produced during infection. UL13 is a serine threonine protein kinase conserved among many herpesviruses. HSV-1 UL13 phosphorylates US3 and thereby controls UL31 and UL34 nuclear rim localization, indicating a role in nuclear egress. Here, we report that HSV-2 UL13 alone induced conformational changes in lamins A and C and redistributed lamin B1 from the nuclear rim to intranuclear granular structures. HSV-2 UL13 directly phosphorylated lamins A, C, and B1 in vitro, and the lamin A1 tail domain. HSV-2 infection recapitulated the lamin alterations seen upon expression of UL13 alone, and other alterations were also observed, indicating that additional viral and/or cellular proteins cooperate with UL13 to alter lamins during HSV-2 infection to allow nuclear egress.

  14. Lamin A/C deficiency causes defective nuclear mechanics and mechanotransduction.

    PubMed

    Lammerding, Jan; Schulze, P Christian; Takahashi, Tomosaburo; Kozlov, Serguei; Sullivan, Teresa; Kamm, Roger D; Stewart, Colin L; Lee, Richard T

    2004-02-01

    Mutations in the lamin A/C gene (LMNA) cause a variety of human diseases including Emery-Dreifuss muscular dystrophy, dilated cardiomyopathy, and Hutchinson-Gilford progeria syndrome. The tissue-specific effects of lamin mutations are unclear, in part because the function of lamin A/C is incompletely defined, but the many muscle-specific phenotypes suggest that defective lamin A/C could increase cellular mechanical sensitivity. To investigate the role of lamin A/C in mechanotransduction, we subjected lamin A/C-deficient mouse embryo fibroblasts to mechanical strain and measured nuclear mechanical properties and strain-induced signaling. We found that Lmna-/- cells have increased nuclear deformation, defective mechanotransduction, and impaired viability under mechanical strain. NF-kappaB-regulated transcription in response to mechanical or cytokine stimulation was attenuated in Lmna-/- cells despite increased transcription factor binding. Lamin A/C deficiency is thus associated with both defective nuclear mechanics and impaired mechanically activated gene transcription. These findings suggest that the tissue-specific effects of lamin A/C mutations observed in the laminopathies may arise from varying degrees of impaired nuclear mechanics and transcriptional activation.

  15. Fast regulation of AP-1 activity through interaction of lamin A/C, ERK1/2, and c-Fos at the nuclear envelope

    PubMed Central

    González, José María; Navarro-Puche, Ana; Casar, Berta; Crespo, Piero; Andrés, Vicente

    2008-01-01

    Sequestration of c-Fos at the nuclear envelope (NE) through interaction with A-type lamins suppresses AP-1–dependent transcription. We show here that c-Fos accumulation within the extraction-resistant nuclear fraction (ERNF) and its interaction with lamin A are reduced and enhanced by gain-of and loss-of ERK1/2 activity, respectively. Moreover, hindering ERK1/2-dependent phosphorylation of c-Fos attenuates its release from the ERNF induced by serum and promotes its interaction with lamin A. Accordingly, serum stimulation rapidly releases preexisting c-Fos from the NE via ERK1/2-dependent phosphorylation, leading to a fast activation of AP-1 before de novo c-Fos synthesis. Moreover, lamin A–null cells exhibit increased AP-1 activity and reduced levels of c-Fos phosphorylation. We also find that active ERK1/2 interacts with lamin A and colocalizes with c-Fos and A-type lamins at the NE. Thus, NE-bound ERK1/2 functions as a molecular switch for rapid mitogen-dependent AP-1 activation through phosphorylation-induced release of preexisting c-Fos from its inhibitory interaction with lamin A/C. PMID:19015316

  16. Structural organization of nuclear lamins A, C, B1, and B2 revealed by superresolution microscopy

    PubMed Central

    Shimi, Takeshi; Kittisopikul, Mark; Tran, Joseph; Goldman, Anne E.; Adam, Stephen A.; Zheng, Yixian; Jaqaman, Khuloud; Goldman, Robert D.

    2015-01-01

    The nuclear lamina is a key structural element of the metazoan nucleus. However, the structural organization of the major proteins composing the lamina is poorly defined. Using three-dimensional structured illumination microscopy and computational image analysis, we characterized the supramolecular structures of lamin A, C, B1, and B2 in mouse embryo fibroblast nuclei. Each isoform forms a distinct fiber meshwork, with comparable physical characteristics with respect to mesh edge length, mesh face area and shape, and edge connectivity to form faces. Some differences were found in face areas among isoforms due to variation in the edge lengths and number of edges per face, suggesting that each meshwork has somewhat unique assembly characteristics. In fibroblasts null for the expression of either lamins A/C or lamin B1, the remaining lamin meshworks are altered compared with the lamin meshworks in wild-type nuclei or nuclei lacking lamin B2. Nuclei lacking LA/C exhibit slightly enlarged meshwork faces and some shape changes, whereas LB1-deficient nuclei exhibit primarily a substantial increase in face area. These studies demonstrate that individual lamin isoforms assemble into complex networks within the nuclear lamina and that A- and B-type lamins have distinct roles in maintaining the organization of the nuclear lamina. PMID:26310440

  17. Lamin Mutations Accelerate Aging via Defective Export of Mitochondrial mRNAs through Nuclear Envelope Budding.

    PubMed

    Li, Yihang; Hassinger, Linda; Thomson, Travis; Ding, Baojin; Ashley, James; Hassinger, William; Budnik, Vivian

    2016-08-08

    Defective RNA metabolism and transport are implicated in aging and degeneration [1, 2], but the underlying mechanisms remain poorly understood. A prevalent feature of aging is mitochondrial deterioration [3]. Here, we link a novel mechanism for RNA export through nuclear envelope (NE) budding [4, 5] that requires A-type lamin, an inner nuclear membrane-associated protein, to accelerated aging observed in Drosophila LaminC (LamC) mutations. These LamC mutations were modeled after A-lamin (LMNA) mutations causing progeroid syndromes (PSs) in humans. We identified mitochondrial assembly regulatory factor (Marf), a mitochondrial fusion factor (mitofusin), as well as other transcripts required for mitochondrial integrity and function, in a screen for RNAs that exit the nucleus through NE budding. PS-modeled LamC mutations induced premature aging in adult flight muscles, including decreased levels of specific mitochondrial protein transcripts (RNA) and progressive mitochondrial degradation. PS-modeled LamC mutations also induced the accelerated appearance of other phenotypes associated with aging, including a progressive accumulation of polyubiquitin aggregates [6, 7] and myofibril disorganization [8, 9]. Consistent with these observations, the mutants had progressive jumping and flight defects. Downregulating marf alone induced the above aging defects. Nevertheless, restoring marf was insufficient for rescuing the aging phenotypes in PS-modeled LamC mutations, as other mitochondrial RNAs are affected by inhibition of NE budding. Analysis of NE budding in dominant and recessive PS-modeled LamC mutations suggests a mechanism by which abnormal lamina organization prevents the egress of these RNAs via NE budding. These studies connect defects in RNA export through NE budding to progressive loss of mitochondrial integrity and premature aging. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Nuclear lamins are not required for lamina-associated domain organization in mouse embryonic stem cells

    PubMed Central

    Amendola, Mario; van Steensel, Bas

    2015-01-01

    In mammals, the nuclear lamina interacts with hundreds of large genomic regions, termed lamina-associated domains (LADs) that are generally in a transcriptionally repressed state. Lamins form the major structural component of the lamina and have been reported to bind DNA and chromatin. Here, we systematically evaluate whether lamins are necessary for the LAD organization in murine embryonic stem cells. Surprisingly, removal of essentially all lamins does not have any detectable effect on the genome-wide interaction pattern of chromatin with emerin, a marker of the inner nuclear membrane. This suggests that other components of the lamina mediate these interactions. PMID:25784758

  19. Nuclear lamins and progerin are dispensable for antioxidant Nrf2 response to arsenic and cadmium.

    PubMed

    Hashimoto, Kazunori; Majumdar, Rima; Tsuji, Yoshiaki

    2017-05-01

    Lamins are important constituents of the nuclear inner membrane and provide a platform for transcription factors and chromatin. Progerin, a C-terminal truncated lamin A mutant, causes premature aging termed Hutchinson-Gilford Progeria Syndrome (HGPS). Oxidative stress appears to be involved in the pathogenesis of HGPS, although the mechanistic role of progerin remains elusive. Here we examined whether nuclear lamins are important for a cellular antioxidant mechanism, and whether progerin compromises it. We investigated the activation of nuclear factor-E2-related factor 2 (Nrf2) which regulates various antioxidant genes including heme oxygenase-1 (HMOX1), following exposure to sodium arsenite or cadmium chloride in lamin knockdown human cell lines and primary HGPS human fibroblasts. Knocking down lamin A/C, or B, or all nuclear lamins simultaneously in three human cell lines (HaCaT, SW480, and K562) did not impair arsenite- or cadmium-induced activation of Nrf2. Progerin-expressing human primary HGPS fibroblasts showed lower basal levels of HMOX1 and NQO1 expression; however, in response to arsenic stress both normal and HGPS primary fibroblasts showed Nrf2 nuclear accumulation along with upregulation and phosphorylation of p62/SQSTM1 at Ser351, downregulation of Keap1, and comparable expression of an array of downstream Nrf2-regulated antioxidant genes. We also observed new forms of cleaved lamin A, B1 and B2 induced by cadmium stress although their roles in the Nrf2 antioxidant system need further investigation. These results suggest that the nuclear lamins and progerin have marginal roles in the activation of the antioxidant Nrf2 response to arsenic and cadmium.

  20. An Absence of Nuclear Lamins in Keratinocytes Leads to Ichthyosis, Defective Epidermal Barrier Function, and Intrusion of Nuclear Membranes and Endoplasmic Reticulum into the Nuclear Chromatin

    PubMed Central

    Jung, Hea-Jin; Tatar, Angelica; Tu, Yiping; Nobumori, Chika; Yang, Shao H.; Goulbourne, Chris N.; Herrmann, Harald

    2014-01-01

    B-type lamins (lamins B1 and B2) have been considered to be essential for many crucial functions in the cell nucleus (e.g., DNA replication and mitotic spindle formation). However, this view has been challenged by the observation that an absence of both B-type lamins in keratinocytes had no effect on cell proliferation or the development of skin and hair. The latter findings raised the possibility that the functions of B-type lamins are subserved by lamins A and C. To explore that idea, we created mice lacking all nuclear lamins in keratinocytes. Those mice developed ichthyosis and a skin barrier defect, which led to death from dehydration within a few days after birth. Microscopy of nuclear-lamin-deficient skin revealed hyperkeratosis and a disordered stratum corneum with an accumulation of neutral lipid droplets; however, BrdU incorporation into keratinocytes was normal. Skin grafting experiments confirmed the stratum corneum abnormalities and normal BrdU uptake. Interestingly, the absence of nuclear lamins in keratinocytes resulted in an interspersion of nuclear/endoplasmic reticulum membranes with the chromatin. Thus, a key function of the nuclear lamina is to serve as a “fence” and prevent the incursion of cytoplasmic organelles into the nuclear chromatin. PMID:25312645

  1. Lamin B1 overexpression increases nuclear rigidity in autosomal dominant leukodystrophy fibroblasts

    PubMed Central

    Ferrera, Denise; Canale, Claudio; Marotta, Roberto; Mazzaro, Nadia; Gritti, Marta; Mazzanti, Michele; Capellari, Sabina; Cortelli, Pietro; Gasparini, Laura

    2014-01-01

    The architecture and structural mechanics of the cell nucleus are defined by the nuclear lamina, which is formed by A- and B-type lamins. Recently, gene duplication and protein overexpression of lamin B1 (LB1) have been reported in pedigrees with autosomal dominant leukodystrophy (ADLD). However, how the overexpression of LB1 affects nuclear mechanics and function and how it may result in pathology remain unexplored. Here, we report that in primary human skin fibroblasts derived from ADLD patients, LB1, but not other lamins, is overexpressed at the nuclear lamina and specifically enhances nuclear stiffness. Transient transfection of LB1 in HEK293 and neuronal N2a cells mimics the mechanical phenotype of ADLD nuclei. Notably, in ADLD fibroblasts, reducing LB1 protein levels by shRNA knockdown restores elasticity values to those indistinguishable from control fibroblasts. Moreover, isolated nuclei from ADLD fibroblasts display a reduced nuclear ion channel open probability on voltage-step application, suggesting that biophysical changes induced by LB1 overexpression may alter nuclear signaling cascades in somatic cells. Overall, the overexpression of LB1 in ADLD cells alters nuclear mechanics and is linked to changes in nuclear signaling, which could help explain the pathogenesis of this disease.—Ferrera, D., Canale, C., Marotta, R., Mazzaro, N., Gritti, M., Mazzanti, M., Capellari, S., Cortelli, P., Gasparini, L. Lamin B1 overexpression increases nuclear rigidity in autosomal dominant leukodystrophy fibroblasts. PMID:24858279

  2. Loss of lamin A function increases chromatin dynamics in the nuclear interior

    NASA Astrophysics Data System (ADS)

    Bronshtein, I.; Kepten, E.; Kanter, I.; Berezin, S.; Lindner, M.; Redwood, Abena B.; Mai, S.; Gonzalo, S.; Foisner, R.; Shav-Tal, Y.; Garini, Y.

    2015-08-01

    Chromatin is organized in a highly ordered yet dynamic manner in the cell nucleus, but the principles governing this organization remain unclear. Similarly, it is unknown whether, and how, various proteins regulate chromatin motion and as a result influence nuclear organization. Here by studying the dynamics of different genomic regions in the nucleus of live cells, we show that the genome has highly constrained dynamics. Interestingly, depletion of lamin A strikingly alters genome dynamics, inducing a dramatic transition from slow anomalous diffusion to fast and normal diffusion. In contrast, depletion of LAP2α, a protein that interacts with lamin A and chromatin, has no such effect on genome dynamics. We speculate that chromosomal inter-chain interactions formed by lamin A throughout the nucleus contribute to chromatin dynamics, and suggest that the molecular regulation of chromatin diffusion by lamin A in the nuclear interior is critical for the maintenance of genome organization.

  3. The highly conserved nuclear lamin Ig-fold binds to PCNA: its role in DNA replication

    PubMed Central

    Shumaker, Dale K.; Solimando, Liliana; Sengupta, Kaushik; Shimi, Takeshi; Adam, Stephen A.; Grunwald, Antje; Strelkov, Sergei V.; Aebi, Ueli; Cardoso, M. Cristina; Goldman, Robert D.

    2008-01-01

    This study provides insights into the role of nuclear lamins in DNA replication. Our data demonstrate that the Ig-fold motif located in the lamin C terminus binds directly to proliferating cell nuclear antigen (PCNA), the processivity factor necessary for the chain elongation phase of DNA replication. We find that the introduction of a mutation in the Ig-fold, which alters its structure and causes human muscular dystrophy, inhibits PCNA binding. Studies of nuclear assembly and DNA replication show that lamins, PCNA, and chromatin are closely associated in situ. Exposure of replicating nuclei to an excess of the lamin domain containing the Ig-fold inhibits DNA replication in a concentration-dependent fashion. This inhibitory effect is significantly diminished in nuclei exposed to the same domain bearing the Ig-fold mutation. Using the crystal structures of the lamin Ig-fold and PCNA, molecular docking simulations suggest probable interaction sites. These findings also provide insights into the mechanisms underlying the numerous disease-causing mutations located within the lamin Ig-fold. PMID:18426975

  4. Differential Predictive Roles of A- and B-Type Nuclear Lamins in Prostate Cancer Progression

    PubMed Central

    Saarinen, Irena; Mirtti, Tuomas; Seikkula, Heikki; Boström, Peter J.; Taimen, Pekka

    2015-01-01

    Background Prostate cancer (PCa) is the most common cancer among men in western countries. While active surveillance is increasingly utilized, the majority of patients are currently treated with radical prostatectomy. In order to avoid over-treatment, there is an indisputable need for reliable biomarkers to identify the potentially aggressive and lethal cases. Nuclear intermediate filament proteins called lamins play a role in chromatin organization, gene expression and cell stiffness. The expression of lamin A is associated with poor outcome in colorectal cancer but to date the prognostic value of the lamins has not been tested in other solid tumors. Methods We studied the expression of different lamins with immunohistochemistry in a tissue microarray material of 501 PCa patients undergoing radical prostatectomy and lymph node dissection. Patients were divided into two staining categories (low and high expression). The correlation of lamin expression with clinicopathological variables was tested and the association of lamin status with biochemical recurrence (BCR) and disease specific survival (DSS) was further analyzed. Results Low expression of lamin A associated with lymph node positivity (p<0.01) but not with other clinicopathological variables and low expression had a borderline independent significant association with DSS (HR = 0.4; 95% CI 0.2–1.0; p = 0.052). Similarly, low lamin C expression associated with poorer survival (HR = 0.2; 95% CI 0.1–0.6; p = 0.004). Lamin B1 expression did not associate with clinicopathological variables but high expression independently predicted BCR in multivariable Cox regression analysis (HR = 1.8; 95% CI 1.1–2.9; p = 0.023). Low expression of lamin B2 correlated with lymph node positivity (p<0.01) and predicted unfavorable DSS (HR = 0.4; 95% CI 0.2–1.0; p = 0.047). Conclusions These results suggest differential roles for lamins in PCa progression. Reduced amounts of lamin A/C and B2 increase risk for lymph node

  5. Differential Predictive Roles of A- and B-Type Nuclear Lamins in Prostate Cancer Progression.

    PubMed

    Saarinen, Irena; Mirtti, Tuomas; Seikkula, Heikki; Boström, Peter J; Taimen, Pekka

    2015-01-01

    Prostate cancer (PCa) is the most common cancer among men in western countries. While active surveillance is increasingly utilized, the majority of patients are currently treated with radical prostatectomy. In order to avoid over-treatment, there is an indisputable need for reliable biomarkers to identify the potentially aggressive and lethal cases. Nuclear intermediate filament proteins called lamins play a role in chromatin organization, gene expression and cell stiffness. The expression of lamin A is associated with poor outcome in colorectal cancer but to date the prognostic value of the lamins has not been tested in other solid tumors. We studied the expression of different lamins with immunohistochemistry in a tissue microarray material of 501 PCa patients undergoing radical prostatectomy and lymph node dissection. Patients were divided into two staining categories (low and high expression). The correlation of lamin expression with clinicopathological variables was tested and the association of lamin status with biochemical recurrence (BCR) and disease specific survival (DSS) was further analyzed. Low expression of lamin A associated with lymph node positivity (p<0.01) but not with other clinicopathological variables and low expression had a borderline independent significant association with DSS (HR = 0.4; 95% CI 0.2-1.0; p = 0.052). Similarly, low lamin C expression associated with poorer survival (HR = 0.2; 95% CI 0.1-0.6; p = 0.004). Lamin B1 expression did not associate with clinicopathological variables but high expression independently predicted BCR in multivariable Cox regression analysis (HR = 1.8; 95% CI 1.1-2.9; p = 0.023). Low expression of lamin B2 correlated with lymph node positivity (p<0.01) and predicted unfavorable DSS (HR = 0.4; 95% CI 0.2-1.0; p = 0.047). These results suggest differential roles for lamins in PCa progression. Reduced amounts of lamin A/C and B2 increase risk for lymph node metastasis and disease specific death possibly

  6. Nuclear networking.

    PubMed

    Xie, Wei; Burke, Brian

    2017-07-04

    Nuclear lamins are intermediate filament proteins that represent important structural components of metazoan nuclear envelopes (NEs). By combining proteomics and superresolution microscopy, we recently reported that both A- and B-type nuclear lamins form spatially distinct filament networks at the nuclear periphery of mouse fibroblasts. In particular, A-type lamins exhibit differential association with nuclear pore complexes (NPCs). Our studies reveal that the nuclear lamina network in mammalian somatic cells is less ordered and more complex than that of amphibian oocytes, the only other system in which the lamina has been visualized at high resolution. In addition, the NPC component Tpr likely links NPCs to the A-type lamin network, an association that appears to be regulated by C-terminal modification of various A-type lamin isoforms. Many questions remain, however, concerning the structure and assembly of lamin filaments, as well as with their mode of association with other nuclear components such as peripheral chromatin.

  7. Organization of the lamin scaffold in the internal nuclear matrix of normal and transformed hepatocytes.

    PubMed

    Barboro, Paola; D'Arrigo, Cristina; Repaci, Erica; Patrone, Eligio; Balbi, Cecilia

    2010-04-01

    Nuclear lamins are among the more abundant proteins making up the internal nuclear matrix, but very little is known about their structure in the nucleoplasm. Using immunoelectron microscopy, we demonstrate the organization of lamins in the nuclear matrix isolated from rat hepatocytes for the first time. Lamin epitopes are arrayed both in locally ordered clusters and in quasi-regular rows. Fourier filtering of the images demonstrates that the epitopes are placed at the nodes and halfway between the nodes of square or rhombic lattices that are about 50 nm on each side, as well as along rows at regular approximately 25-nm intervals. In addition, we have compared this structure with that of the internal nuclear matrix isolated from persistent hepatocyte nodules. In transformed hepatocytes, the islands of lamin lattice are lost, and only a partial regularity in the rows of gold particles remains. We suggest that orthogonal lattice assembly might be an intrinsic property of lamin molecules, and that the disassembly may be triggered by simple molecular events such as phosphorylation. Copyright (c) 2009 Elsevier Inc. All rights reserved.

  8. Nuclear lamin stiffness is a barrier to 3D migration, but softness can limit survival.

    PubMed

    Harada, Takamasa; Swift, Joe; Irianto, Jerome; Shin, Jae-Won; Spinler, Kyle R; Athirasala, Avathamsa; Diegmiller, Rocky; Dingal, P C Dave P; Ivanovska, Irena L; Discher, Dennis E

    2014-03-03

    Cell migration through solid tissue often involves large contortions of the nucleus, but biological significance is largely unclear. The nucleoskeletal protein lamin-A varies both within and between cell types and was shown here to contribute to cell sorting and survival in migration through constraining micropores. Lamin-A proved rate-limiting in 3D migration of diverse human cells that ranged from glioma and adenocarcinoma lines to primary mesenchymal stem cells (MSCs). Stoichiometry of A- to B-type lamins established an activation barrier, with high lamin-A:B producing extruded nuclear shapes after migration. Because the juxtaposed A and B polymer assemblies respectively conferred viscous and elastic stiffness to the nucleus, subpopulations with different A:B levels sorted in 3D migration. However, net migration was also biphasic in lamin-A, as wild-type lamin-A levels protected against stress-induced death, whereas deep knockdown caused broad defects in stress resistance. In vivo xenografts proved consistent with A:B-based cell sorting, and intermediate A:B-enhanced tumor growth. Lamins thus impede 3D migration but also promote survival against migration-induced stresses.

  9. Invertebrate lamins

    SciTech Connect

    Melcer, Shai; Gruenbaum, Yosef . E-mail: gru@vms.huji.ac.il; Krohne, Georg . E-mail: krohne@biozentrum.uni-wuerzburg.de

    2007-06-10

    Lamins are the main component of the nuclear lamina and considered to be the ancestors of all intermediate filament proteins. They are localized mainly at the nuclear periphery where they form protein complexes with integral proteins of the nuclear inner membrane, transcriptional regulators, histones and chromatin modifiers. Studying lamins in invertebrate species has unique advantages including the smaller number of lamin genes in the invertebrate genomes and powerful genetic analyses in Caenorhabditis elegans and Drosophila melanogaster. These simpler nuclear lamina systems allow direct analyses of their structure and functions. Here we give an overview of recent advances in the field of invertebrate nuclear lamins with special emphasis on their evolution, assembly and functions.

  10. B-type nuclear lamin and the nuclear pore complex Nup107-160 influences maintenance of the spindle envelope required for cytokinesis in Drosophila male meiosis

    PubMed Central

    Hayashi, Daisuke; Tanabe, Karin; Katsube, Hiroka

    2016-01-01

    ABSTRACT In higher eukaryotes, nuclear envelope (NE) disassembly allows chromatin to condense and spindle microtubules to access kinetochores. The nuclear lamina, which strengthens the NE, is composed of a polymer meshwork made of A- and B-type lamins. We found that the B-type lamin (Lam) is not fully disassembled and continues to localize along the spindle envelope structure during Drosophila male meiosis I, while the A-type lamin (LamC) is completely dispersed throughout the cytoplasm. Among the nuclear pore complex proteins, Nup107 co-localized with Lam during this meiotic division. Surprisingly, Lam depletion resulted in a higher frequency of cytokinesis failure in male meiosis. We also observed the similar meiotic phenotype in Nup107-depleted cells. Abnormal localization of Lam was found in the Nup-depleted cells at premeiotic and meiotic stages. The central spindle microtubules became abnormal and recruitment of a contractile ring component to the cleavage sites was disrupted in Lam-depleted cells and Nup107-depleted cells. Therefore, we speculate that both proteins are required for a reinforcement of the spindle envelope, which supports the formation of central spindle microtubules essential for cytokinesis in Drosophila male meiosis. PMID:27402967

  11. Granulocytic nuclear differentiation of lamin B receptor-deficient mouse EPRO cells.

    PubMed

    Zwerger, Monika; Herrmann, Harald; Gaines, Peter; Olins, Ada L; Olins, Donald E

    2008-08-01

    Lamin B receptor (LBR) is an integral protein of the inner nuclear membrane. Recent studies have demonstrated that genetic deficiency of LBR during granulopoiesis results in hypolobulation of the mature neutrophil nucleus, as observed in human Pelger-Huët anomaly and mouse ichthyosis (ic). In this study, we utilized differentiated early promyelocytes (EPRO cells) that were derived from the bone marrow of homozygous and heterozygous ichthyosis mice to examine changes to the expression of nuclear envelope proteins and heterochromatin structure that result from deficient LBR expression. Wild-type (+/+), heterozygous (+/ic), and homozygous (ic/ic) granulocytic forms of EPRO cells were analyzed for the expression of multiple lamins and inner nuclear envelope proteins by immunostaining and immunoblotting techniques. The heterochromatin architecture was also examined by immunostaining for histone lysine methylation. Wild-type (+/+) and heterozygous (+/ic) granulocytic forms revealed ring-shaped nuclei and contained LBR within the nuclear envelope; ic/ic granulocytes exhibited smaller ovoid nuclei devoid of LBR. The pericentric heterochromatin of undifferentiated and granulocytic ic/ic cells was condensed into larger spots and shifted away from the nuclear envelope, compared to +/+ and +/ic cell forms. Lamin A/C, which is normally not present in mature granulocytes, was significantly elevated in LBR-deficient EPRO cells. Our observations suggest roles for LBR during granulopoiesis, which can involve augmenting nuclear membrane growth, facilitating compartmentalization of heterochromatin, and promoting downregulation of lamin A/C expression.

  12. Defects in nuclear structure and function promote dilated cardiomyopathy in lamin A/C–deficient mice

    PubMed Central

    Nikolova, Vesna; Leimena, Christiana; McMahon, Aisling C.; Tan, Ju Chiat; Chandar, Suchitra; Jogia, Dilesh; Kesteven, Scott H.; Michalicek, Jan; Otway, Robyn; Verheyen, Fons; Rainer, Stephen; Stewart, Colin L.; Martin, David; Feneley, Michael P.; Fatkin, Diane

    2004-01-01

    Laminopathies are a group of disorders caused by mutations in the LMNA gene that encodes the nuclear lamina proteins, lamin A and lamin C; their pathophysiological basis is unknown. We report that lamin A/C–deficient (Lmna–/–) mice develop rapidly progressive dilated cardiomyopathy (DCM) characterized by left ventricular (LV) dilation and reduced systolic contraction. Isolated Lmna–/– myocytes show reduced shortening with normal baseline and peak amplitude of Ca2+ transients. Lmna–/– LV myocyte nuclei have marked alterations of shape and size with central displacement and fragmentation of heterochromatin; these changes are present but less severe in left atrial nuclei. Electron microscopy of Lmna–/– cardiomyocytes shows disorganization and detachment of desmin filaments from the nuclear surface with progressive disruption of the cytoskeletal desmin network. Alterations in nuclear architecture are associated with defective nuclear function evidenced by decreased SREBP1 import, reduced PPARγ expression, and a lack of hypertrophic gene activation. These findings suggest a model in which the primary pathophysiological mechanism in Lmna–/– mice is defective force transmission resulting from disruption of lamin interactions with the muscle-specific desmin network and loss of cytoskeletal tension. Despite severe DCM, defects in nuclear function prevent Lmna–/– cardiomyocytes from developing compensatory hypertrophy and accelerate disease progression. PMID:14755333

  13. Differential Expression of A-Type and B-Type Lamins during Hair Cycling

    PubMed Central

    Hanif, Mubashir; Rozell, Björn; Eriksson, Maria

    2009-01-01

    Multiple genetic disorders caused by mutations that affect the proteins lamin A and C show strong skin phenotypes. These disorders include the premature aging disorders Hutchinson-Gilford progeria syndrome and mandibuloacral dysplasia, as well as restrictive dermopathy. Prior studies have shown that the lamin A/C and B proteins are expressed in skin, but little is known about their normal expression in the different skin cell-types and during the hair cycle. Our immunohistochemical staining for lamins A/C and B in wild-type mice revealed strong expression in the basal cell layer of the epidermis, the outer root sheath, and the dermal papilla during all stages of the hair cycle. Lower expression of both lamins A/C and B was seen in suprabasal cells of the epidermis, in the hypodermis, and in the bulb of catagen follicles. In addition, we have utilized a previously described mouse model of Hutchinson-Gilford progeria syndrome and show here that the expression of progerin does not result in pronounced effects on hair cycling or the expression of lamin B. PMID:19122810

  14. Lamin-dependent Localization of UNC-84, A Protein Required for Nuclear Migration in Caenorhabditis elegans

    PubMed Central

    Lee, Kenneth K.; Starr, Daniel; Cohen, Merav; Liu, Jun; Han, Min; Wilson, Katherine L.; Gruenbaum, Yosef

    2002-01-01

    Mutations in the Caenorhabditis elegans unc-84 gene cause defects in nuclear migration and anchoring. We show that endogenous UNC-84 protein colocalizes with Ce-lamin at the nuclear envelope and that the envelope localization of UNC-84 requires Ce-lamin. We also show that during mitosis, UNC-84 remains at the nuclear periphery until late anaphase, similar to known inner nuclear membrane proteins. UNC-84 protein is first detected at the 26-cell stage and thereafter is present in most cells during development and in adults. UNC-84 is properly expressed in unc-83 and anc-1 lines, which have phenotypes similar to unc-84, suggesting that neither the expression nor nuclear envelope localization of UNC-84 depends on UNC-83 or ANC-1 proteins. The envelope localization of Ce-lamin, Ce-emerin, Ce-MAN1, and nucleoporins are unaffected by the loss of UNC-84. UNC-84 is not required for centrosome attachment to the nucleus because centrosomes are localized normally in unc-84 hyp7 cells despite a nuclear migration defect. Models for UNC-84 localization are discussed. PMID:11907270

  15. AFM single-cell force spectroscopy links altered nuclear and cytoskeletal mechanics to defective cell adhesion in cardiac myocytes with a nuclear lamin mutation

    PubMed Central

    Lanzicher, Thomas; Martinelli, Valentina; Long, Carlin S; Del Favero, Giorgia; Puzzi, Luca; Borelli, Massimo; Mestroni, Luisa; Taylor, Matthew R G; Sbaizero, Orfeo

    2015-01-01

    Previous investigations suggested that lamin A/C gene (LMNA) mutations, which cause a variety of human diseases including muscular dystrophies and cardiomyopathies, alter the nuclear mechanical properties. We hypothesized that biomechanical changes may extend beyond the nucleus. PMID:26309016

  16. In silico synchronization reveals regulators of nuclear ruptures in lamin A/C deficient model cells

    PubMed Central

    Robijns, J.; Molenberghs, F.; Sieprath, T.; Corne, T. D. J.; Verschuuren, M.; De Vos, W. H.

    2016-01-01

    The nuclear lamina is a critical regulator of nuclear structure and function. Nuclei from laminopathy patient cells experience repetitive disruptions of the nuclear envelope, causing transient intermingling of nuclear and cytoplasmic components. The exact causes and consequences of these events are not fully understood, but their stochastic occurrence complicates in-depth analyses. To resolve this, we have established a method that enables quantitative investigation of spontaneous nuclear ruptures, based on co-expression of a firmly bound nuclear reference marker and a fluorescent protein that shuttles between the nucleus and cytoplasm during ruptures. Minimally invasive imaging of both reporters, combined with automated tracking and in silico synchronization of individual rupture events, allowed extracting information on rupture frequency and recovery kinetics. Using this approach, we found that rupture frequency correlates inversely with lamin A/C levels, and can be reduced in genome-edited LMNA knockout cells by blocking actomyosin contractility or inhibiting the acetyl-transferase protein NAT10. Nuclear signal recovery followed a kinetic that is co-determined by the severity of the rupture event, and could be prolonged by knockdown of the ESCRT-III complex component CHMP4B. In conclusion, our approach reveals regulators of nuclear rupture induction and repair, which may have critical roles in disease development. PMID:27461848

  17. In silico synchronization reveals regulators of nuclear ruptures in lamin A/C deficient model cells

    NASA Astrophysics Data System (ADS)

    Robijns, J.; Molenberghs, F.; Sieprath, T.; Corne, T. D. J.; Verschuuren, M.; de Vos, W. H.

    2016-07-01

    The nuclear lamina is a critical regulator of nuclear structure and function. Nuclei from laminopathy patient cells experience repetitive disruptions of the nuclear envelope, causing transient intermingling of nuclear and cytoplasmic components. The exact causes and consequences of these events are not fully understood, but their stochastic occurrence complicates in-depth analyses. To resolve this, we have established a method that enables quantitative investigation of spontaneous nuclear ruptures, based on co-expression of a firmly bound nuclear reference marker and a fluorescent protein that shuttles between the nucleus and cytoplasm during ruptures. Minimally invasive imaging of both reporters, combined with automated tracking and in silico synchronization of individual rupture events, allowed extracting information on rupture frequency and recovery kinetics. Using this approach, we found that rupture frequency correlates inversely with lamin A/C levels, and can be reduced in genome-edited LMNA knockout cells by blocking actomyosin contractility or inhibiting the acetyl-transferase protein NAT10. Nuclear signal recovery followed a kinetic that is co-determined by the severity of the rupture event, and could be prolonged by knockdown of the ESCRT-III complex component CHMP4B. In conclusion, our approach reveals regulators of nuclear rupture induction and repair, which may have critical roles in disease development.

  18. β-Amyloid induces nuclear protease-mediated lamin fragmentation independent of caspase activation.

    PubMed

    Ramasamy, Vijay Sankar; Islam, Md Imamul; Haque, Md Aminul; Shin, Song Yub; Park, Il-Seon

    2016-06-01

    β-Amyloid (Aβ), a hallmark peptide of Alzheimer's disease, induces both caspase-dependent apoptosis and non-apoptotic cell death. In this study, we examined caspase-independent non-apoptotic cell death preceding caspase activation in Aβ42-treated cells. We first determined the optimal treatment conditions for inducing cell death without caspase activation and selected a double-treatment method involving the incubation of cells with Aβ42 for 4 and 6 h (4+6 h sample). We observed that levels of lamin A (LA) and lamin B (LB) were reduced in the 4+6 h samples. This reduction was decreased by treatment with suc-AAPF-CMK, an inhibitor of nuclear scaffold (NS) protease, but not by treatment with z-VAD-FMK, a pan-caspase inhibitor. In addition, suc-AAPF-CMK decreased the changes in nuclear morphology observed in cells in the 4+6 h samples, which were different from nuclear fragmentation observed in STS-treated cells. Furthermore, suc-AAPF-CMK inhibited cell death in the 4+6 h samples. LA and LB fragmentation occurred in the isolated nuclei and was also inhibited by suc-AAPF-CMK. Together, these data indicated that the fragmentation of LA and LB in the Aβ42-treated cells was induced by an NS protease, whose identity is not clearly determined yet. A correlation between Aβ42 toxicity and the lamin fragmentation by NS protease suggests that inhibition of the protease could be an effective method for controlling the pathological process of AD.

  19. Nuclear export signal-interacting protein forms complexes with lamin A/C-Nups to mediate the CRM1-independent nuclear export of large hepatitis delta antigen.

    PubMed

    Huang, Cheng; Jiang, Jia-Yin; Chang, Shin C; Tsay, Yeou-Guang; Chen, Mei-Ru; Chang, Ming-Fu

    2013-02-01

    Nuclear export is an important process that not only regulates the functions of cellular factors but also facilitates the assembly of viral nucleoprotein complexes. Chromosome region maintenance 1 (CRM1) that mediates the transport of proteins bearing the classical leucine-rich nuclear export signal (NES) is the best-characterized nuclear export receptor. Recently, several CRM1-independent nuclear export pathways were also identified. The nuclear export of the large form of hepatitis delta antigen (HDAg-L), a nucleocapsid protein of hepatitis delta virus (HDV), which contains a CRM1-independent proline-rich NES, is mediated by the host NES-interacting protein (NESI). The mechanism of the NESI protein in mediating nuclear export is still unknown. In this study, NESI was characterized as a highly glycosylated membrane protein. It interacted and colocalized well in the nuclear envelope with lamin A/C and nucleoporins. Importantly, HDAg-L could be coimmunoprecipitated with lamin A/C and nucleoporins. In addition, binding of the cargo HDAg-L to the C terminus of NESI was detected for the wild-type protein but not for the nuclear export-defective HDAg-L carrying a P205A mutation [HDAg-L(P205A)]. Knockdown of lamin A/C effectively reduced the nuclear export of HDAg-L and the assembly of HDV. These data indicate that by forming complexes with lamin A/C and nucleoporins, NESI facilitates the CRM1-independent nuclear export of HDAg-L.

  20. Accumulation of mutant lamin A causes progressive changes in nuclear architecture in Hutchinson-Gilford progeria syndrome.

    PubMed

    Goldman, Robert D; Shumaker, Dale K; Erdos, Michael R; Eriksson, Maria; Goldman, Anne E; Gordon, Leslie B; Gruenbaum, Yosef; Khuon, Satya; Mendez, Melissa; Varga, Renée; Collins, Francis S

    2004-06-15

    Hutchinson-Gilford progeria syndrome (HGPS) is a premature aging disorder, commonly caused by a point mutation in the lamin A gene that results in a protein lacking 50 aa near the C terminus, denoted LADelta50. Here we show by light and electron microscopy that HGPS is associated with significant changes in nuclear shape, including lobulation of the nuclear envelope, thickening of the nuclear lamina, loss of peripheral heterochromatin, and clustering of nuclear pores. These structural defects worsen as HGPS cells age in culture, and their severity correlates with an apparent increase in LADelta50. Introduction of LADelta50 into normal cells by transfection or protein injection induces the same changes. We hypothesize that these alterations in nuclear structure are due to a concentration-dependent dominant-negative effect of LADelta50, leading to the disruption of lamin-related functions ranging from the maintenance of nuclear shape to regulation of gene expression and DNA replication.

  1. AKT1-mediated Lamin A/C degradation is required for nuclear degradation and normal epidermal terminal differentiation.

    PubMed

    Naeem, A S; Zhu, Y; Di, W L; Marmiroli, S; O'Shaughnessy, R F L

    2015-12-01

    Nuclear degradation is a key stage in keratinocyte terminal differentiation and the formation of the cornified envelope that comprises the majority of epidermal barrier function. Parakeratosis, the retention of nuclear material in the cornified layer of the epidermis, is a common histological observation in many skin diseases, notably in atopic dermatitis and psoriasis. Keratinocyte nuclear degradation is not well characterised, and it is unclear whether the retained nuclei contribute to the altered epidermal differentiation seen in eczema and psoriasis. Loss of AKT1 function strongly correlated with parakeratosis both in eczema samples and in organotypic culture models. Although levels of DNAses, including DNase1L2, were unchanged, proteomic analysis revealed an increase in Lamin A/C. AKT phosphorylates Lamin A/C, targeting it for degradation. Consistent with this, Lamin A/C degradation was inhibited and Lamin A/C was observed in the cornified layer of AKT1 knockdown organotypic cultures, surrounding retained nuclear material. Using AKT-phosphorylation-dead Lamin A constructs we show that the retention of nuclear material is sufficient to cause profound changes in epidermal terminal differentiation, specifically a reduction in Loricrin, Keratin 1, Keratin 10, and filaggrin expression. We show that preventing nuclear degradation upregulates BMP2 expression and SMAD1 signalling. Consistent with these data, we observe both parakeratosis and evidence of increased SMAD1 signalling in atopic dermatitis. We therefore present a model that, in the absence of AKT1-mediated Lamin A/C degradation, DNA degradation processes, such as those mediated by DNAse 1L2, are prevented, leading to parakeratosis and changes in epidermal differentiation.

  2. C. elegans Nuclear Envelope Proteins Emerin, MAN1, Lamin, and Nucleoporins Reveal Unique Timing of Nuclear Envelope Breakdown during Mitosis

    PubMed Central

    Lee, Kenneth K.; Gruenbaum, Yosef; Spann, Perah; Liu, Jun; Wilson, Katherine L.

    2000-01-01

    Emerin, MAN1, and LAP2 are integral membrane proteins of the vertebrate nuclear envelope. They share a 43-residue N-terminal motif termed the LEM domain. We found three putative LEM domain genes in Caenorhabditis elegans, designated emr-1, lem-2, and lem-3. We analyzed emr-l, which encodes Ce-emerin, and lem-2, which encodes Ce-MAN1. Ce-emerin and Ce-MAN1 migrate on SDS-PAGE as 17- and 52-kDa proteins, respectively. Based on their biochemical extraction properties and immunolocalization, both Ce-emerin and Ce-MAN1 are integral membrane proteins localized at the nuclear envelope. We used antibodies against Ce-MAN1, Ce-emerin, nucleoporins, and Ce-lamin to determine the timing of nuclear envelope breakdown during mitosis in C. elegans. The C. elegans nuclear envelope disassembles very late compared with vertebrates and Drosophila. The nuclear membranes remained intact everywhere except near spindle poles during metaphase and early anaphase, fully disassembling only during mid-late anaphase. Disassembly of pore complexes, and to a lesser extent the lamina, depended on embryo age: pore complexes were absent during metaphase in >30-cell embryos but existed until anaphase in 2- to 24-cell embryos. Intranuclear mRNA splicing factors disassembled after prophase. The timing of nuclear disassembly in C. elegans is novel and may reflect its evolutionary position between unicellular and more complex eukaryotes. PMID:10982402

  3. Dysregulated interactions between lamin A and SUN1 induce abnormalities in the nuclear envelope and endoplasmic reticulum in progeric laminopathies.

    PubMed

    Chen, Zi-Jie; Wang, Wan-Ping; Chen, Yu-Ching; Wang, Jing-Ya; Lin, Wen-Hsin; Tai, Lin-Ai; Liou, Gan-Guang; Yang, Chung-Shi; Chi, Ya-Hui

    2014-04-15

    Hutchinson-Gilford progeria syndrome (HGPS) is a human progeroid disease caused by a point mutation on the LMNA gene. We reported previously that the accumulation of the nuclear envelope protein SUN1 contributes to HGPS nuclear aberrancies. However, the mechanism by which interactions between mutant lamin A (also known as progerin or LAΔ50) and SUN1 produce HGPS cellular phenotypes requires further elucidation. Using light and electron microscopy, this study demonstrated that SUN1 contributes to progerin-elicited structural changes in the nuclear envelope and the endoplasmic reticulum (ER) network. We further identified two domains through which full-length lamin A associates with SUN1, and determined that the farnesylated cysteine within the CaaX motif of lamin A has a stronger affinity for SUN1 than does the lamin A region containing amino acids 607 to 656. Farnesylation of progerin enhanced its interaction with SUN1 and reduced SUN1 mobility, thereby promoting the aberrant recruitment of progerin to the ER membrane during postmitotic assembly of the nuclear envelope, resulting in the accumulation of SUN1 over consecutive cellular divisions. These results indicate that the dysregulated interaction of SUN1 and progerin in the ER during nuclear envelope reformation determines the progression of HGPS.

  4. Detection of Endogenous Nuclear Proteins in Plant Cells: Localizing Nuclear Matrix Constituent Proteins (NMCPs), the Plant Analogs of Lamins.

    PubMed

    Ciska, Malgorzata; de la Espina, Susana Moreno Díaz

    2017-01-01

    At present, two complementary approaches are used for in situ protein visualization in plant nuclei. Imaging of transformed fluorescent proteins is the election tool for the analysis of protein movement and interaction. However, this methodology presents several drawbacks for the identification/localization of endogenous nuclear factors, such as over-expression or mislocalization of transformed proteins. In contrast, immunocytochemistry with specific antibodies represents a powerful tool for the localization of endogenous nuclear proteins at their "native" nuclear sub-compartments. In plant cells, the cell wall hampers antibody accessibility during immunocytochemical analysis thereby reducing the effectivity of the technique, particularly in the case of lowly expressed proteins. To overcome this problem in nuclear protein immunodetection, we developed a method based on the in vitro incubation of isolated nuclei with specific antibodies followed by imaging by confocal fluorescence or electron microscopy. Here we describe the application of this methodology to the localization of Nuclear Matrix Constituent Proteins (NMCP), the plant analogs of lamins, of the monocot Allium cepa, using antibodies raised against highly conserved regions of the proteins.

  5. Photoexcited calphostin C selectively destroys nuclear lamin B1 in neoplastic human and rat cells - a novel mechanism of action of a photodynamic tumor therapy agent.

    PubMed

    Chiarini, Anna; Whitfield, James F; Pacchiana, Raffaella; Armato, Ubaldo; Dal Pra, Ilaria

    2008-09-01

    Lamin B1, a major component of the nuclear lamina, anchors the nucleus to the cytoskeletal cage, and controls nuclear orientation, chromosome positioning and, alongside several enzymes, fundamental nuclear functions. Exposing polyomavirus-transformed rat pyF111 fibroblasts and human cervical carcinoma (HCC) C4-I cells for 30 min to photoexcited perylenequinone calphostin C, i.e. Cal C(phiE), an established reactive oxygen species (ROS)-generator and protein kinase C (PKC) inhibitor, caused the cells to selectively oxidize and then totally destroy their nuclear lamin B1 by only 60 min after starting the treatment, i.e. when apoptotic caspases' activities had not yet increased. However, while the oxidized lamin B1 was being destroyed, lamins A/C, the lamin A-associated nuclear envelope protein emerin, and the nucleoplasmic protein cyclin E were neither oxidized nor destroyed. The oxidized lamin B was ubiquitinated and demolished in the proteasome probably by an enhanced peptidyl-glutaminase-like activity. Hence, the Cal C(phiE)-induced rapid and selective lamin B1 oxidation and proteasomal destruction ahead of the activation of apoptotic caspases was by itself a most severe molecular lesion impairing vital nuclear functions. Conversely, Cal C directly added to the cells kept in the dark damaged neither nuclear lamin B1 nor cell viability. Thus, our findings reveal a novel cell-damaging mechanism of a photodynamic tumor therapeutic agent.

  6. Phosphorylation of lamins determine their structural properties and signaling functions

    PubMed Central

    Torvaldson, Elin; Kochin, Vitaly; Eriksson, John E

    2015-01-01

    Lamin A/C is part of the nuclear lamina, a meshwork of intermediate filaments underlying the inner nuclear membrane. The lamin network is anchoring a complex set of structural and linker proteins and is either directly or through partner proteins also associated or interacting with a number of signaling protein and transcription factors. During mitosis the nuclear lamina is dissociated by well established phosphorylation- dependent mechanisms. A-type lamins are, however, also phosphorylated during interphase. A recent study identified 20 interphase phosphorylation sites on lamin A/C and explored their functions related to lamin dynamics; movements, localization and solubility. Here we discuss these findings in the light of lamin functions in health and disease. PMID:25793944

  7. Polyphyly of nuclear lamin genes indicates an early eukaryotic origin of the metazoan-type intermediate filament proteins.

    PubMed

    Kollmar, Martin

    2015-05-29

    The nuclear lamina is a protein meshwork associated with the inner side of the nuclear envelope contributing structural, signalling and regulatory functions. Here, I report on the evolution of an important component of the lamina, the lamin intermediate filament proteins, across the eukaryotic tree of life. The lamins show a variety of protein domain and sequence motif architectures beyond the classical α-helical rod, nuclear localisation signal, immunoglobulin domain and CaaX motif organisation, suggesting extension and adaptation of functions in many species. I identified lamin genes not only in metazoa and Amoebozoa as previously described, but also in other opisthokonts including Ichthyosporea and choanoflagellates, in oomycetes, a sub-family of Stramenopiles, and in Rhizaria, implying that they must have been present very early in eukaryotic evolution if not even the last common ancestor of all extant eukaryotes. These data considerably extend the current perception of lamin evolution and have important implications with regard to the evolution of the nuclear envelope.

  8. The effect of the lamin A and its mutants on nuclear structure, cell proliferation, protein stability, and mobility in embryonic cells.

    PubMed

    Piekarowicz, Katarzyna; Machowska, Magdalena; Dratkiewicz, Ewelina; Lorek, Daria; Madej-Pilarczyk, Agnieszka; Rzepecki, Ryszard

    2017-08-01

    LMNA gene encodes for nuclear intermediate filament proteins lamin A/C. Mutations in this gene lead to a spectrum of genetic disorders, collectively referred to as laminopathies. Lamin A/C are widely expressed in most differentiated somatic cells but not in early embryos and some undifferentiated cells. To investigate the role of lamin A/C in cell phenotype maintenance and differentiation, which could be a determinant of the pathogenesis of laminopathies, we examined the role played by exogenous lamin A and its mutants in differentiated cell lines (HeLa, NHDF) and less-differentiated HEK 293 cells. We introduced exogenous wild-type and mutated (H222P, L263P, E358K D446V, and ∆50) lamin A into different cell types and analyzed proteins' impact on proliferation, protein mobility, and endogenous nuclear envelope protein distribution. The mutants give rise to a broad spectrum of nuclear phenotypes and relocate lamin C. The mutations ∆50 and D446V enhance proliferation in comparison to wild-type lamin A and control cells, but no changes in exogenous protein mobility measured by FRAP were observed. Interestingly, although transcripts for lamins A and C are at similar level in HEK 293 cells, only lamin C protein is detected in western blots. Also, exogenous lamin A and its mutants, when expressed in HEK 293 cells underwent posttranscriptional processing. Overall, our results provide new insight into the maintenance of lamin A in less-differentiated cells. Embryonic cells are very sensitive to lamin A imbalance, and its upregulation disturbs lamin C, which may influence gene expression and many regulatory pathways.

  9. The Different Function of Single Phosphorylation Sites of Drosophila melanogaster Lamin Dm and Lamin C

    PubMed Central

    Zaremba-Czogalla, Magdalena; Piekarowicz, Katarzyna; Wachowicz, Katarzyna; Kozioł, Katarzyna; Dubińska-Magiera, Magda; Rzepecki, Ryszard

    2012-01-01

    Lamins' functions are regulated by phosphorylation at specific sites but our understanding of the role of such modifications is practically limited to the function of cdc 2 (cdk1) kinase sites in depolymerization of the nuclear lamina during mitosis. In our study we used Drosophila lamin Dm (B-type) to examine the function of particular phosphorylation sites using pseudophosphorylated mutants mimicking single phosphorylation at experimentally confirmed in vivo phosphosites (S25E, S45E, T435E, S595E). We also analyzed lamin C (A-type) and its mutant S37E representing the N-terminal cdc2 (mitotic) site as well as lamin Dm R64H mutant as a control, non-polymerizing lamin. In the polymerization assay we could observe different effects of N-terminal cdc2 site pseudophosphorylation on A- and B-type lamins: lamin Dm S45E mutant was insoluble, in contrast to lamin C S37E. Lamin Dm T435E (C-terminal cdc2 site) and R64H were soluble in vitro. We also confirmed that none of the single phosphorylation site modifications affected the chromatin binding of lamin Dm, in contrast to the lamin C N-terminal cdc2 site. In vivo, all lamin Dm mutants were incorporated efficiently into the nuclear lamina in transfected Drosophila S2 and HeLa cells, although significant amounts of S45E and T435E were also located in cytoplasm. When farnesylation incompetent mutants were expressed in HeLa cells, lamin Dm T435E was cytoplasmic and showed higher mobility in FRAP assay. PMID:22393432

  10. The role of nuclear lamin B1 in cell proliferation and senescence

    PubMed Central

    Shimi, Takeshi; Butin-Israeli, Veronika; Adam, Stephen A.; Hamanaka, Robert B.; Goldman, Anne E.; Lucas, Catherine A.; Shumaker, Dale K.; Kosak, Steven T.; Chandel, Navdeep S.; Goldman, Robert D.

    2011-01-01

    Nuclear lamin B1 (LB1) is a major structural component of the nucleus that appears to be involved in the regulation of many nuclear functions. The results of this study demonstrate that LB1 expression in WI-38 cells decreases during cellular senescence. Premature senescence induced by oncogenic Ras also decreases LB1 expression through a retinoblastoma protein (pRb)-dependent mechanism. Silencing the expression of LB1 slows cell proliferation and induces premature senescence in WI-38 cells. The effects of LB1 silencing on proliferation require the activation of p53, but not pRb. However, the induction of premature senescence requires both p53 and pRb. The proliferation defects induced by silencing LB1 are accompanied by a p53-dependent reduction in mitochondrial reactive oxygen species (ROS), which can be rescued by growth under hypoxic conditions. In contrast to the effects of LB1 silencing, overexpression of LB1 increases the proliferation rate and delays the onset of senescence of WI-38 cells. This overexpression eventually leads to cell cycle arrest at the G1/S boundary. These results demonstrate the importance of LB1 in regulating the proliferation and senescence of human diploid cells through a ROS signaling pathway. PMID:22155925

  11. Werner complex deficiency in cells disrupts the Nuclear Pore Complex and the distribution of lamin B1.

    PubMed

    Li, Zhi; Zhu, Yizhou; Zhai, Yujia; R Castroagudin, Michelle; Bao, Yifei; White, Tommy E; Glavy, Joseph S

    2013-12-01

    From the surrounding shell to the inner machinery, nuclear proteins provide the functional plasticity of the nucleus. This study highlights the nuclear association of Pore membrane (POM) protein NDC1 and Werner protein (WRN), a RecQ helicase responsible for the DNA instability progeria disorder, Werner Syndrome. In our previous publication, we connected the DNA damage sensor Werner's Helicase Interacting Protein (WHIP), a binding partner of WRN, to the NPC. Here, we confirm the association of the WRN/WHIP complex and NDC1. In established WRN/WHIP knockout cell lines, we further demonstrate the interdependence of WRN/WHIP and Nucleoporins (Nups). These changes do not completely abrogate the barrier of the Nuclear Envelope (NE) but do affect the distribution of FG Nups and the RAN gradient, which are necessary for nuclear transport. Evidence from WRN/WHIP knockout cell lines demonstrates changes in the processing and nucleolar localization of lamin B1. The appearance of "RAN holes" void of RAN corresponds to regions within the nucleolus filled with condensed pools of lamin B1. From WRN/WHIP knockout cell line extracts, we found three forms of lamin B1 that correspond to mature holoprotein and two potential post-translationally modified forms of the protein. Upon treatment with topoisomerase inhibitors lamin B1 cleavage occurs only in WRN/WHIP knockout cells. Our data suggest the link of the NDC1 and WRN as one facet of the network between the nuclear periphery and genome stability. Loss of WRN complex leads to multiple alterations at the NPC and the nucleolus. © 2013. Published by Elsevier B.V. All rights reserved.

  12. Low lamin A expression in lung adenocarcinoma cells from pleural effusions is a pejorative factor associated with high number of metastatic sites and poor Performance status.

    PubMed

    Kaspi, Elise; Frankel, Diane; Guinde, Julien; Perrin, Sophie; Laroumagne, Sophie; Robaglia-Schlupp, Andrée; Ostacolo, Kevin; Harhouri, Karim; Tazi-Mezalek, Rachid; Micallef, Joelle; Dutau, Hervé; Tomasini, Pascale; De Sandre-Giovannoli, Annachiara; Lévy, Nicolas; Cau, Pierre; Astoul, Philippe; Roll, Patrice

    2017-01-01

    The type V intermediate filament lamins are the principal components of the nuclear matrix, including the nuclear lamina. Lamins are divided into A-type and B-type, which are encoded by three genes, LMNA, LMNB1, and LMNB2. The alternative splicing of LMNA produces two major A-type lamins, lamin A and lamin C. Previous studies have suggested that lamins are involved in cancer development and progression. A-type lamins have been proposed as biomarkers for cancer diagnosis, prognosis, and/or follow-up. The aim of the present study was to investigate lamins in cancer cells from metastatic pleural effusions using immunofluorescence, western blotting, and flow cytometry. In a sub-group of lung adenocarcinomas, we found reduced expression of lamin A but not of lamin C. The reduction in lamin A expression was correlated with the loss of epithelial membrane antigen (EMA)/MUC-1, an epithelial marker that is involved in the epithelial to mesenchymal transition (EMT). Finally, the lamin A expression was inversely correlated with the number of metastatic sites and the WHO Performance status, and association of pleural, bone and lung metastatic localizations was more frequent when lamin A expression was reduced. In conclusion, low lamin A but not lamin C expression in pleural metastatic cells could represent a major actor in the development of metastasis, associated with EMT and could account for a pejorative factor correlated with a poor Performance status.

  13. Src1 is a Protein of the Inner Nuclear Membrane Interacting with the Dictyostelium Lamin NE81.

    PubMed

    Batsios, Petros; Ren, Xiang; Baumann, Otto; Larochelle, Denis A; Gräf, Ralph

    2016-03-18

    The nuclear envelope (NE) consists of the outer and inner nuclear membrane (INM), whereby the latter is bound to the nuclear lamina. Src1 is a Dictyostelium homologue of the helix-extension-helix family of proteins, which also includes the human lamin-binding protein MAN1. Both endogenous Src1 and GFP-Src1 are localized to the NE during the entire cell cycle. Immuno-electron microscopy and light microscopy after differential detergent treatment indicated that Src1 resides in the INM. FRAP experiments with GFP-Src1 cells suggested that at least a fraction of the protein could be stably engaged in forming the nuclear lamina together with the Dictyostelium lamin NE81. Both a BioID proximity assay and mis-localization of soluble, truncated mRFP-Src1 at cytosolic clusters consisting of an intentionally mis-localized mutant of GFP-NE81 confirmed an interaction of Src1 and NE81. Expression GFP-Src1(1-646), a fragment C-terminally truncated after the first transmembrane domain, disrupted interaction of nuclear membranes with the nuclear lamina, as cells formed protrusions of the NE that were dependent on cytoskeletal pulling forces. Protrusions were dependent on intact microtubules but not actin filaments. Our results indicate that Src1 is required for integrity of the NE and highlight Dictyostelium as a promising model for the evolution of nuclear architecture.

  14. Nuclear processes associated with peculiar A-type stars.

    NASA Technical Reports Server (NTRS)

    Cameron, A. G. W.

    1971-01-01

    A discussion is given of the various roles that nuclear reactions may play in production of anomalous abundances of elements in peculiar A stars. The effects which may be expected to occur both in the surface nuclear reactions and in some possible internal reactions that can occur in advanced stages of stellar evolution are considered. It is suggested that various features of peculiar A stars may require simultaneous operation of two or more of the processes of surface diffusion, surface nuclear reactions, and internal nuclear reactions.

  15. Chrom3D: three-dimensional genome modeling from Hi-C and nuclear lamin-genome contacts.

    PubMed

    Paulsen, Jonas; Sekelja, Monika; Oldenburg, Anja R; Barateau, Alice; Briand, Nolwenn; Delbarre, Erwan; Shah, Akshay; Sørensen, Anita L; Vigouroux, Corinne; Buendia, Brigitte; Collas, Philippe

    2017-01-30

    Current three-dimensional (3D) genome modeling platforms are limited by their inability to account for radial placement of loci in the nucleus. We present Chrom3D, a user-friendly whole-genome 3D computational modeling framework that simulates positions of topologically-associated domains (TADs) relative to each other and to the nuclear periphery. Chrom3D integrates chromosome conformation capture (Hi-C) and lamin-associated domain (LAD) datasets to generate structure ensembles that recapitulate radial distributions of TADs detected in single cells. Chrom3D reveals unexpected spatial features of LAD regulation in cells from patients with a laminopathy-causing lamin mutation. Chrom3D is freely available on github.

  16. Lamin A and lamin C form homodimers and coexist in higher complex forms both in the nucleoplasmic fraction and in the lamina of cultured human cells.

    PubMed

    Kolb, Thorsten; Maass, Kendra; Hergt, Michaela; Aebi, Ueli; Herrmann, Harald

    2011-01-01

    We have investigated and quantified the nuclear A-type lamin pool from human HeLa S3 suspension cells with respect to their distribution to detergent soluble and insoluble fractions. We devised a sequential extraction protocol and found that maximally 10% of A-type lamins are recovered in the soluble fraction. Notably, lamin C is enriched in low detergent fractions and only with 0.5% Nonidet P-40 lamin A and C are recovered in ratios nearly equivalent to those found in whole cell extracts and in the lamina fraction. Authentic nucleoplasmic proteins such as LAP2a, pRB and p53 are co-extracted to a large part together with the A-type lamins in these fractions. By sucrose density centrifugation we revealed that the majority of lamins co-sedimented with human IgG indicating they form rather small complexes in the range of dimers and slightly larger complexes. Some lamin A - but not lamin C - is obtained in addition in a much faster sedimenting fraction. Authentic nuclear proteins such as PCNA, p53 and LAP2a were found both in the light and the heavy sucrose fractions together with lamin A. Last but not least, immunoprecipitation experiments from both soluble fractions and from RIPA lysates of whole cells revealed that lamin A and lamin C do not form heterodimers but segregate practically completely. Correspondingly, immunofluorescence microscopy of formaldehyde-fixed cells clearly demonstrated that lamin A and C are localized at least in part to distinct patches within the lamina. Hence, the structural segregation of lamin A and C is indeed retained in the nuclear envelope to some extent too.

  17. The major nucleoside triphosphatase in pea (Pisum sativum L.) nuclei and in rat liver nuclei share common epitopes also present in nuclear lamins

    NASA Technical Reports Server (NTRS)

    Tong, C. G.; Dauwalder, M.; Clawson, G. A.; Hatem, C. L.; Roux, S. J.

    1993-01-01

    The major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum L.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. The major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) G2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes it in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb G2 in mammalian nuclei. The pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by G2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase is probably derived from a lamin precursor by proteolysis.

  18. The major nucleoside triphosphatase in pea (Pisum sativum L.) nuclei and in rat liver nuclei share common epitopes also present in nuclear lamins

    NASA Technical Reports Server (NTRS)

    Tong, C. G.; Dauwalder, M.; Clawson, G. A.; Hatem, C. L.; Roux, S. J.

    1993-01-01

    The major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum L.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. The major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) G2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes it in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb G2 in mammalian nuclei. The pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by G2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase is probably derived from a lamin precursor by proteolysis.

  19. From the nucleus to the plasma membrane: translocation of the nuclear proteins histone H3 and lamin B1 in apoptotic microglia.

    PubMed

    Klein, Barbara; Lütz-Meindl, Ursula; Kerschbaum, Hubert H

    2014-05-01

    Nuclear autoantibodies have been found in patients with autoimmune diseases. One possible source for nuclear antigens are apoptotic cells. However, the mechanism of how apoptotic cells make nuclear factors accessible to the immune system is still elusive. In the present study, we investigated the redistribution of nuclear components after UV irradiation in the microglial cell line BV-2 and in primary mouse microglia at the ultrastructural level. We used transmission electron microscopy-coupled electron energy loss spectroscopy (EELS) to measure phosphorus as an indicator for nucleic acids and immunogold labeling to detect histone H3 and lamin B1 in apoptotic cells. EELS revealed elevated concentrations of phosphorus in nuclear and cytoplasmic condensed chromatin compared to the remaining cytoplasm. Furthermore, immunolabeling of lamin B1 and histone H3 was detected in apoptotic microglia not only in the nucleus, but also in the cytoplasm, and even at the plasma membrane. Confocal images of apoptotic microglia, which were not previously permeabilized, showed patches of histone H3 and lamin B1 labeling at the cell surface. The pan-caspase inhibitor Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone) prevented the occurrence of cytoplasmic condensed chromatin in apoptotic microglia. Our findings indicate that nuclear components leak from the nucleus into the cytoplasm in apoptotic microglia. At least histone H3 and lamin B1 reach the cell surface, this may promote autoreactive processes.

  20. Simple Separation of Functionally Distinct Populations of Lamin-Binding Proteins.

    PubMed

    Berk, Jason M; Wilson, Katherine L

    2016-01-01

    The inner membrane of the nuclear envelope (NE) is home to hundreds of integral membrane proteins (NE transmembrane proteins, "NETs") with conserved or tissue-specific roles in genome organization and nuclear function. Nearly all characterized NETs bind A- or B-type lamins directly. However, hundreds of NETs remain uncharacterized, collectively posing an enormous gap that must be bridged to understand nuclear function and genome biology. We provide technically simple protocols for the separation and recovery of functionally distinct populations of NETs and A-type lamins. This protocol was developed for emerin, an inner nuclear membrane protein that binds lamins and barrier-to-autointegration factor (BANF1) as a component of nuclear lamina structure, and has diverse roles in nuclear assembly, signaling, and gene regulation. This protocol separates easily solubilized ("easy") populations of nuclear lamina proteins (emerin, lamin A, BAF) from "sonication-dependent" populations. Depending on cell type, the "easy" and "sonication-dependent" fractions each contain up to about half the available emerin, A-type lamins, and BAF, whereas B-type lamins and histone H3 are predominantly sonication dependent. The two populations of emerin have distinct posttranslational modifications, and only one population associates with BAF. This method may be useful for functional screening or analysis of other lamin-associated proteins, including novel NETs emerging from proteomic studies.

  1. Calphostin C, a remarkable multimodal photodynamic killer of neoplastic cells by selective nuclear lamin B1 destruction and apoptogenesis (Review).

    PubMed

    Chiarini, Anna; Whitfield, James F; Pacchiana, Raffaella; Marconi, Maddalena; Armato, Ubaldo; Dal Prà, Ilaria

    2010-04-01

    Perylenequinones that generate reactive oxygen species (ROS) when illuminated with visible light have been recommended as photodynamic chemotherapeutic agents. One of these is calphostin C (CalC), the action of the photo-activated derivative of which, CalCphiE, has been ascribed to its ability to selectively and irreversibly inhibit protein kinase Cs (PKCs). But recent results of experiments with neoplastic rat fibroblasts and human breast and uterine cervix cancer cells have revealed that the action of CalCphiE involves more than PKC inhibition. Besides suppressing PKC activity, CalCphiE rapidly causes endoplasmic reticulum (ER) stress in breast cancer cells and the selective complete oxidation and proteasomal destruction of the functionally essential nuclear envelope protein lamin B1, in human cervical carcinoma (HCC) cells and neoplastic rat fibroblasts. When these lamin B1-lacking cells are placed in the dark, cytoplasmic membrane-linked PKC activities suddenly rebound and apoptogenesis is initiated as indicated by the immediate release of cytochrome c from mitochondria and later on the activation of caspases. Hence, CalCphiE is a photodynamic cytocidal agent attacking multiple targets in cancer cells and it would be worth determining, even for their best applicative use, whether other perylenequinones also share the so far unexpectedly complex deadly properties of the CalCphiE.

  2. Myopathic Lamin Mutations Cause Reductive Stress and Activate the Nrf2/Keap-1 Pathway

    PubMed Central

    Dialynas, George; Shrestha, Om K.; Ponce, Jessica M.; Zwerger, Monika; Thiemann, Dylan A.; Young, Grant H.; Moore, Steven A.; Yu, Liping; Lammerding, Jan; Wallrath, Lori L.

    2015-01-01

    Mutations in the human LMNA gene cause muscular dystrophy by mechanisms that are incompletely understood. The LMNA gene encodes A-type lamins, intermediate filaments that form a network underlying the inner nuclear membrane, providing structural support for the nucleus and organizing the genome. To better understand the pathogenesis caused by mutant lamins, we performed a structural and functional analysis on LMNA missense mutations identified in muscular dystrophy patients. These mutations perturb the tertiary structure of the conserved A-type lamin Ig-fold domain. To identify the effects of these structural perturbations on lamin function, we modeled these mutations in Drosophila Lamin C and expressed the mutant lamins in muscle. We found that the structural perturbations had minimal dominant effects on nuclear stiffness, suggesting that the muscle pathology was not accompanied by major structural disruption of the peripheral nuclear lamina. However, subtle alterations in the lamina network and subnuclear reorganization of lamins remain possible. Affected muscles had cytoplasmic aggregation of lamins and additional nuclear envelope proteins. Transcription profiling revealed upregulation of many Nrf2 target genes. Nrf2 is normally sequestered in the cytoplasm by Keap-1. Under oxidative stress Nrf2 dissociates from Keap-1, translocates into the nucleus, and activates gene expression. Unexpectedly, biochemical analyses revealed high levels of reducing agents, indicative of reductive stress. The accumulation of cytoplasmic lamin aggregates correlated with elevated levels of the autophagy adaptor p62/SQSTM1, which also binds Keap-1, abrogating Nrf2 cytoplasmic sequestration, allowing Nrf2 nuclear translocation and target gene activation. Elevated p62/SQSTM1 and nuclear enrichment of Nrf2 were identified in muscle biopsies from the corresponding muscular dystrophy patients, validating the disease relevance of our Drosophila model. Thus, novel connections were made

  3. Incomplete processing of mutant lamin A in Hutchinson-Gilford progeria leads to nuclear abnormalities, which are reversed by farnesyltransferase inhibition.

    PubMed

    Glynn, Michael W; Glover, Thomas W

    2005-10-15

    Hutchinson-Gilford progeria syndrome (HGPS) is typically caused by mutations in codon 608 (G608G) of the LMNA gene, which activates a cryptic splice site resulting in the in-frame loss of 150 nucleotides from the lamin A message. The deleted region includes a protein cleavage site that normally removes 15 amino acids, including a CAAX box farnesylation site, from the lamin A protein. We investigated the processing of the C-terminus of the mutant protein, 'progerin', and found that it does not undergo cleavage and, indeed, remains farnesylated. The retention of the farnesyl group may have numerous consequences, as farnesyl groups increase lipophilicity and are involved in membrane association and in protein interactions, and is likely to be an important factor in the HGPS phenotype. To further investigate this, we studied the effects of farnesylation inhibition on nuclear phenotypes in cells expressing normal and mutant lamin A. Expression of a GFP-progerin fusion protein in normal fibroblasts caused a high incidence of nuclear abnormalities, as was also seen in HGPS fibroblasts, and resulted in abnormal nuclear localization of GFP-progerin in comparison with the localization pattern of GFP-lamin A. Expression of a GFP-lamin A fusion containing a mutation preventing the final cleavage step, causing the protein to remain farnesylated, displayed identical localization patterns and nuclear abnormalities as in HGPS cells and in cells expressing GFP-progerin. Exposure to a farnesyltransferase inhibitor (FTI), PD169541, caused a significant improvement in the nuclear morphology of cells expressing GFP-progerin and in HGPS cells. These results implicate the abnormal farnesylation of progerin in the cellular phenotype in HGPS cells and suggest that FTIs may represent a therapeutic option for patients with HGPS.

  4. Recent advances in understanding the role of lamins in health and disease

    PubMed Central

    Reddy, Sita; Comai, Lucio

    2016-01-01

    Lamins are major components of the nuclear lamina, a network of proteins that supports the nuclear envelope in metazoan cells. Over the past decade, biochemical studies have provided support for the view that lamins are not passive bystanders providing mechanical stability to the nucleus but play an active role in the organization of the genome and the function of fundamental nuclear processes. It has also become apparent that lamins are critical for human health, as a large number of mutations identified in the gene that encodes for A-type lamins are associated with tissue-specific and systemic genetic diseases, including the accelerated aging disorder known as Hutchinson-Gilford progeria syndrome. Recent years have witnessed great advances in our understanding of the role of lamins in the nucleus and the functional consequences of disease-associated A-type lamin mutations. Many of these findings have been presented in comprehensive reviews. In this mini-review, we discuss recent breakthroughs in the role of lamins in health and disease and what lies ahead in lamin research. PMID:27803806

  5. Nuclear envelope proteins Nesprin2 and LaminA regulate proliferation and apoptosis of vascular endothelial cells in response to shear stress.

    PubMed

    Han, Yue; Wang, Lu; Yao, Qing-Ping; Zhang, Ping; Liu, Bo; Wang, Guo-Liang; Shen, Bao-Rong; Cheng, Binbin; Wang, Yingxiao; Jiang, Zong-Lai; Qi, Ying-Xin

    2015-05-01

    The dysfunction of vascular endothelial cells (ECs) influenced by flow shear stress is crucial for vascular remodeling. However, the roles of nuclear envelope (NE) proteins in shear stress-induced EC dysfunction are still unknown. Our results indicated that, compared with normal shear stress (NSS), low shear stress (LowSS) suppressed the expression of two types of NE proteins, Nesprin2 and LaminA, and increased the proliferation and apoptosis of ECs. Targeted small interfering RNA (siRNA) and gene overexpression plasmid transfection revealed that Nesprin2 and LaminA participate in the regulation of EC proliferation and apoptosis. A protein/DNA array was further used to detect the activation of transcription factors in ECs following transfection with target siRNAs and overexpression plasmids. The regulation of AP-2 and TFIID mediated by Nesprin2 and the activation of Stat-1, Stat-3, Stat-5 and Stat-6 by LaminA were verified under shear stress. Furthermore, using Ingenuity Pathway Analysis software and real-time RT-PCR, the effects of Nesprin2 or LaminA on the downstream target genes of AP-2, TFIID, and Stat-1, Stat-3, Stat-5 and Stat-6, respectively, were investigated under LowSS. Our study has revealed that NE proteins are novel mechano-sensitive molecules in ECs. LowSS suppresses the expression of Nesprin2 and LaminA, which may subsequently modulate the activation of important transcription factors and eventually lead to EC dysfunction.

  6. Viscoelastic Behavior of Human Lamin A Proteins in the Context of Dilated Cardiomyopathy

    PubMed Central

    Krishnaswamy, Rema; Bhattacharjee, Pritha; Ray, Pulak; Sood, Ajay K.; Sengupta, Kaushik

    2013-01-01

    Lamins are intermediate filament proteins of type V constituting a nuclear lamina or filamentous meshwork which lines the nucleoplasmic side of the inner nuclear membrane. This protein mesh provides a supporting scaffold for the nuclear envelope and tethers interphase chromosome to the nuclear periphery. Mutations of mainly A-type lamins are found to be causative for at least 11 human diseases collectively termed as laminopathies majority of which are characterised by aberrant nuclei with altered structural rigidity, deformability and poor mechanotransduction behaviour. But the investigation of viscoelastic behavior of lamin A continues to elude the field. In order to address this problem, we hereby present the very first report on viscoelastic properties of wild type human lamin A and some of its mutants linked with Dilated cardiomyopathy (DCM) using quantitative rheological measurements. We observed a dramatic strain-softening effect on lamin A network as an outcome of the strain amplitude sweep measurements which could arise from the large compliance of the quasi-cross-links in the network or that of the lamin A rods. In addition, the drastic stiffening of the differential elastic moduli on superposition of rotational and oscillatory shear stress reflect the increase in the stiffness of the laterally associated lamin A rods. These findings present a preliminary insight into distinct biomechanical properties of wild type lamin A protein and its mutants which in turn revealed interesting differences. PMID:24386194

  7. The Caenorhabditis elegans SUN protein UNC-84 interacts with lamin to transfer forces from the cytoplasm to the nucleoskeleton during nuclear migration.

    PubMed

    Bone, Courtney R; Tapley, Erin C; Gorjánácz, Mátyás; Starr, Daniel A

    2014-09-15

    Nuclear migration is a critical component of many cellular and developmental processes. The nuclear envelope forms a barrier between the cytoplasm, where mechanical forces are generated, and the nucleoskeleton. The LINC complex consists of KASH proteins in the outer nuclear membrane and SUN proteins in the inner nuclear membrane that bridge the nuclear envelope. How forces are transferred from the LINC complex to the nucleoskeleton is poorly understood. The Caenorhabditis elegans lamin, LMN-1, is required for nuclear migration and interacts with the nucleoplasmic domain of the SUN protein UNC-84. This interaction is weakened by the unc-84(P91S) missense mutation. These mutant nuclei have an intermediate nuclear migration defect-live imaging of nuclei or LMN-1::GFP shows that many nuclei migrate normally, others initiate migration before subsequently failing, and others fail to begin migration. At least one other component of the nucleoskeleton, the NET5/Samp1/Ima1 homologue SAMP-1, plays a role in nuclear migration. We propose a nut-and-bolt model to explain how forces are dissipated across the nuclear envelope during nuclear migration. In this model, SUN/KASH bridges serve as bolts through the nuclear envelope, and nucleoskeleton components LMN-1 and SAMP-1 act as both nuts and washers on the inside of the nucleus.

  8. The Caenorhabditis elegans SUN protein UNC-84 interacts with lamin to transfer forces from the cytoplasm to the nucleoskeleton during nuclear migration

    PubMed Central

    Bone, Courtney R.; Tapley, Erin C.; Gorjánácz, Mátyás; Starr, Daniel A.

    2014-01-01

    Nuclear migration is a critical component of many cellular and developmental processes. The nuclear envelope forms a barrier between the cytoplasm, where mechanical forces are generated, and the nucleoskeleton. The LINC complex consists of KASH proteins in the outer nuclear membrane and SUN proteins in the inner nuclear membrane that bridge the nuclear envelope. How forces are transferred from the LINC complex to the nucleoskeleton is poorly understood. The Caenorhabditis elegans lamin, LMN-1, is required for nuclear migration and interacts with the nucleoplasmic domain of the SUN protein UNC-84. This interaction is weakened by the unc-84(P91S) missense mutation. These mutant nuclei have an intermediate nuclear migration defect—live imaging of nuclei or LMN-1::GFP shows that many nuclei migrate normally, others initiate migration before subsequently failing, and others fail to begin migration. At least one other component of the nucleoskeleton, the NET5/Samp1/Ima1 homologue SAMP-1, plays a role in nuclear migration. We propose a nut-and-bolt model to explain how forces are dissipated across the nuclear envelope during nuclear migration. In this model, SUN/KASH bridges serve as bolts through the nuclear envelope, and nucleoskeleton components LMN-1 and SAMP-1 act as both nuts and washers on the inside of the nucleus. PMID:25057012

  9. Implications and Assessment of the Elastic Behavior of Lamins in Laminopathies

    PubMed Central

    Dutta, Subarna; Bhattacharyya, Maitree; Sengupta, Kaushik

    2016-01-01

    Lamins are mechanosensitive and elastic components of the nuclear lamina that respond to external mechanical cues by altering gene regulation in a feedback mechanism. Numerous mutations in A-type lamins cause a plethora of diverse diseases collectively termed as laminopathies, the majority of which are characterized by irregularly shaped, fragile, and plastic nuclei. These nuclei are challenged to normal mechanotransduction and lead to disease phenotypes. Here, we review our current understanding of the nucleocytoskeleton coupling in mechanotransduction mediated by lamins. We also present an up-to-date understanding of the methods used to determine laminar elasticity both at the bulk and single molecule level. PMID:27754432

  10. Physiological and Pathological Aging Affects Chromatin Dynamics, Structure and Function at the Nuclear Edge.

    PubMed

    Robin, Jérôme D; Magdinier, Frédérique

    2016-01-01

    Lamins are intermediate filaments that form a complex meshwork at the inner nuclear membrane. Mammalian cells express two types of Lamins, Lamins A/C and Lamins B, encoded by three different genes, LMNA, LMNB1, and LMNB2. Mutations in the LMNA gene are associated with a group of phenotypically diverse diseases referred to as laminopathies. Lamins interact with a large number of binding partners including proteins of the nuclear envelope but also chromatin-associated factors. Lamins not only constitute a scaffold for nuclear shape, rigidity and resistance to stress but also contribute to the organization of chromatin and chromosomal domains. We will discuss here the impact of A-type Lamins loss on alterations of chromatin organization and formation of chromatin domains and how disorganization of the lamina contributes to the patho-physiology of premature aging syndromes.

  11. Physiological and Pathological Aging Affects Chromatin Dynamics, Structure and Function at the Nuclear Edge

    PubMed Central

    Robin, Jérôme D.; Magdinier, Frédérique

    2016-01-01

    Lamins are intermediate filaments that form a complex meshwork at the inner nuclear membrane. Mammalian cells express two types of Lamins, Lamins A/C and Lamins B, encoded by three different genes, LMNA, LMNB1, and LMNB2. Mutations in the LMNA gene are associated with a group of phenotypically diverse diseases referred to as laminopathies. Lamins interact with a large number of binding partners including proteins of the nuclear envelope but also chromatin-associated factors. Lamins not only constitute a scaffold for nuclear shape, rigidity and resistance to stress but also contribute to the organization of chromatin and chromosomal domains. We will discuss here the impact of A-type Lamins loss on alterations of chromatin organization and formation of chromatin domains and how disorganization of the lamina contributes to the patho-physiology of premature aging syndromes. PMID:27602048

  12. Disruption of lamin B1 and lamin B2 processing and localization by farnesyltransferase inhibitors.

    PubMed

    Adam, Stephen A; Butin-Israeli, Veronika; Cleland, Megan M; Shimi, Takeshi; Goldman, Robert D

    2013-01-01

    Lamin A and the B-type lamins, lamin B1 and lamin B2, are translated as pre-proteins that are modified at a carboxyl terminal CAAX motif by farnesylation, proteolysis and carboxymethylation. Lamin A is further processed by proteolysis to remove the farnesyl, but B-type lamins remain permanently farnesylated. Two childhood diseases, Hutchinson Gilford Progeria Syndrome and restrictive dermopathy are caused by defects in the processing of lamin A, resulting in permanent farnesylation of the protein. Farnesyltransferase inhibitors, originally developed to target oncogenic Ras, have recently been used in clinical trials to treat children with Hutchinson Gilford Progeria Syndrome. Lamin B1 and lamin B2 play important roles in cell proliferation and organ development, but little is known about the role of farnesylation in their functions. Treating normal human fibroblasts with farnesyltransferase inhibitors causes the accumulation of unprocessed lamin B2 and lamin A and a decrease in mature lamin B1. Normally, lamins are concentrated at the nuclear envelope/lamina, but when farnesylation is inhibited, the peripheral localization of lamin B2 decreases as its nucleoplasmic levels increase. Unprocessed prelamin A distributes into both the nuclear envelope/lamina and nucleoplasm. Farnesyltransferase inhibitors also cause a rapid cell cycle arrest leading to cellular senescence. This study suggests that the long-term inhibition of protein farnesylation could have unforeseen consequences on nuclear functions.

  13. Significance of host cell kinases in herpes simplex virus type 1 egress and lamin-associated protein disassembly from the nuclear lamina

    SciTech Connect

    Leach, Natalie R.; Roller, Richard J.

    2010-10-10

    The nuclear lamina is thought to be a steric barrier to the herpesvirus capsid. Disruption of the lamina accompanied by phosphorylation of lamina proteins is a conserved feature of herpesvirus infection. In HSV-1-infected cells, protein kinase C (PKC) alpha and delta isoforms are recruited to the nuclear membrane and PKC delta has been implicated in phosphorylation of emerin and lamin B. We tested two critical hypotheses about the mechanism and significance of lamina disruption. First, we show that chemical inhibition of all PKC isoforms reduced viral growth five-fold and inhibited capsid egress from the nucleus. However, specific inhibition of either conventional PKCs or PKC delta does not inhibit viral growth. Second, we show hyperphosphorylation of emerin by viral and cellular kinases is required for its disassociation from the lamina. These data support hypothesis that phosphorylation of lamina components mediates lamina disruption during HSV nuclear egress.

  14. Muscle dystrophy-causing ΔK32 lamin A/C mutant does not impair functions of nucleoplasmic LAP2α - lamin A/C complexes in mice

    PubMed Central

    Pilat, Ursula; Dechat, Thomas; Bertrand, Anne T; Woisetschläger, Nikola; Gotic, Ivana; Spilka, Rita; Biadasiewicz, Katarzyna; Bonne, Gisèle; Foisner, Roland

    2015-01-01

    Summary A-type lamins are components of the nuclear lamina, a filamentous network of the nuclear envelope in metazoans that supports nuclear architecture. In addition, lamin A/C can also be found in the nuclear interior. This nucleoplasmic lamin pool is soluble in physiological buffer, depends on the presence of the lamin-binding protein, Lamina-associated polypeptide 2α (LAP2α) and regulates cell cycle progression in tissue progenitor cells. ΔK32 mutations in A-type lamins cause severe congenital muscle disease in humans and a muscle maturation defect in LmnaΔK32/ΔK32 knock-in mice. At molecular level, mutant ΔK32 lamin A/C protein levels were reduced and all mutant lamin A/C was soluble and mislocalized to the nucleoplasm. To test the role of LAP2α in nucleoplasmic ΔK32 lamin A/C regulation and functions, we deleted LAP2α in LmnaΔK32/ΔK32 knock-in mice. In double mutant mice the LmnaΔK32/ΔK32- linked muscle defect was unaffected. LAP2α interacted with mutant lamin A/C, but unlike wild-type lamin A/C, the intranuclear localization of ΔK32 lamin A/C was not affected by loss of LAP2α. In contrast, loss of LAP2α in LmnaΔK32/ΔK32 mice impaired the regulation of tissue progenitor cells like in lamin A/C wild type animals. These data indicate that a LAP2α-independent assembly defect of ΔK32 lamin A/C is predominant for the mouse pathology, while the LAP2α-linked functions of nucleoplasmic lamin A/C in the regulation of tissue progenitor cells are not affected in LmnaΔK32/ΔK32 mice. PMID:23444379

  15. Specific contribution of lamin A and lamin C in the development of laminopathies

    SciTech Connect

    Sylvius, Nicolas Hathaway, Andrea; Boudreau, Emilie; Gupta, Pallavi; Labib, Sarah; Bolongo, Pierrette M.; Rippstein, Peter; McBride, Heidi; Bilinska, Zofia T.; Tesson, Frederique

    2008-08-01

    Mutations in the lamin A/C gene are involved in multiple human disorders for which the pathophysiological mechanisms are partially understood. Conflicting results prevail regarding the organization of lamin A and C mutants within the nuclear envelope (NE) and on the interactions of each lamin to its counterpart. We over-expressed various lamin A and C mutants both independently and together in COS7 cells. When expressed alone, lamin A with cardiac/muscular disorder mutations forms abnormal aggregates inside the NE and not inside the nucleoplasm. Conversely, the equivalent lamin C organizes as intranucleoplasmic aggregates that never connect to the NE as opposed to wild type lamin C. Interestingly, the lamin C molecules present within these aggregates exhibit an abnormal increased mobility. When co-expressed, the complex formed by lamin A/C aggregates in the NE. Lamin A and C mutants for lipodystrophy behave similarly to the wild type. These findings reveal that lamins A and C may be differentially affected depending on the mutation. This results in multiple possible physiological consequences which likely contribute in the phenotypic variability of laminopathies. The inability of lamin C mutants to join the nuclear rim in the absence of lamin A is a potential pathophysiological mechanism for laminopathies.

  16. Laminated electromagnetic pump stator core

    DOEpatents

    Fanning, A.W.

    1995-08-08

    A stator core for an electromagnetic pump includes a plurality of circumferentially abutting tapered laminations extending radially outwardly from a centerline axis to collectively define a radially inner bore and a radially outer circumference. Each of the laminations includes radially inner and outer edges and has a thickness increasing from the inner edge toward the outer edge to provide a substantially continuous path adjacent the circumference. This pump is used in nuclear fission reactors. 19 figs.

  17. Lamin B1 and lamin B2 are long-lived proteins with distinct functions in retinal development

    PubMed Central

    Razafsky, David; Ward, Candace; Potter, Chloe; Zhu, Wanqiu; Xue, Yunlu; Kefalov, Vladimir J.; Fong, Loren G.; Young, Stephen G.; Hodzic, Didier

    2016-01-01

    Lamin B1 and lamin B2 are essential building blocks of the nuclear lamina, a filamentous meshwork lining the nucleoplasmic side of the inner nuclear membrane. Deficiencies in lamin B1 and lamin B2 impair neurodevelopment, but distinct functions for the two proteins in the development and homeostasis of the CNS have been elusive. Here we show that embryonic depletion of lamin B1 in retinal progenitors and postmitotic neurons affects nuclear integrity, leads to the collapse of the laminB2 meshwork, impairs neuronal survival, and markedly reduces the cellularity of adult retinas. In stark contrast, a deficiency of lamin B2 in the embryonic retina has no obvious effect on lamin B1 localization or nuclear integrity in embryonic retinas, suggesting that lamin B1, but not lamin B2, is strictly required for nucleokinesis during embryonic neurogenesis. However, the absence of lamin B2 prevents proper lamination of adult retinal neurons, impairs synaptogenesis, and reduces cone photoreceptor survival. We also show that lamin B1 and lamin B2 are extremely long-lived proteins in rod and cone photoreceptors. OF interest, a complete absence of both proteins during postnatal life has little or no effect on the survival and function of cone photoreceptors. PMID:27075175

  18. Acute laminitis.

    PubMed

    Baxter, G M

    1994-12-01

    Laminitis is an inflammation of the sensitive laminae along the dorsal aspect of the digit and is considered to be a secondary complication of several predisposing or primary factors. Affected horses are usually very lame, have increased digital pulses, are painful to hoof testers along the toe of the foot, and have evidence of downward rotation or distal displacement of the distal phalanx present on radiographs. Treatments for acute laminitis include anti-inflammatory drugs, anti-endotoxin therapy, vasodilators, antithrombotic therapy, corrective trimming and shoeing, and surgical procedures. Treatment regimens are very controversial and the true efficacy of these treatments is unknown. The quality of laminae damage that occurs with laminitis, however, probably has greater influence on the success of treatment and outcome of the horse than the treatment regimen itself.

  19. Photoresist laminate

    DOEpatents

    Andrade, A.D.; Galbraith, L.K.

    1979-10-01

    The disclosure relates to a laminated negative dry-film photoresist for the production of thick, as well as thin, patterns with vertical sidewalls. Uniform depthwise exposure in a photoresist layer is effected by the use of an ultraviolet filtering top layer.

  20. Structure of the lamin A/C R482W mutant responsible for dominant familial partial lipodystrophy (FPLD)

    SciTech Connect

    Magracheva, Eugenia; Kozlov, Serguei; Stewart, Colin L.; Wlodawer, Alexander; Zdanov, Alexander

    2009-08-07

    Proteins of the A-type lamin family, which consists of two members, lamin A and lamin C, are the major components of a thin proteinaceous filamentous meshwork, the lamina, that underlies the inner nuclear membrane. A-type lamins have recently become the focus of extensive functional studies as a consequence of the linking of at least eight congenital diseases to mutations in the lamin A/C gene (LMNA). This spectrum of pathologies, which mostly manifest themselves as dominant traits, includes muscle dystrophies, dilated cardiomyopathies, the premature aging syndrome Hutchinson-Guilford progeria and familial partial lipodystrophy (FPLD). The crystal structure of the lamin A/C mutant R482W, a variant that causes FPLD, has been determined at 1.5 {angstrom} resolution. A completely novel aggregation state of the C-terminal globular domain and the position of the mutated amino-acid residue suggest means by which the mutation may affect lamin A/C-protein and protein-DNA interactions.

  1. The molecular architecture of lamins in somatic cells.

    PubMed

    Turgay, Yagmur; Eibauer, Matthias; Goldman, Anne E; Shimi, Takeshi; Khayat, Maayan; Ben-Harush, Kfir; Dubrovsky-Gaupp, Anna; Sapra, K Tanuj; Goldman, Robert D; Medalia, Ohad

    2017-03-09

    The nuclear lamina is a fundamental constituent of metazoan nuclei. It is composed mainly of lamins, which are intermediate filament proteins that assemble into a filamentous meshwork, bridging the nuclear envelope and chromatin. Besides providing structural stability to the nucleus, the lamina is involved in many nuclear activities, including chromatin organization, transcription and replication. However, the structural organization of the nuclear lamina is poorly understood. Here we use cryo-electron tomography to obtain a detailed view of the organization of the lamin meshwork within the lamina. Data analysis of individual lamin filaments resolves a globular-decorated fibre appearance and shows that A- and B-type lamins assemble into tetrameric filaments of 3.5 nm thickness. Thus, lamins exhibit a structure that is remarkably different from the other canonical cytoskeletal elements. Our findings define the architecture of the nuclear lamin meshworks at molecular resolution, providing insights into their role in scaffolding the nuclear lamina.

  2. Lamin B receptor

    PubMed Central

    Olins, Ada L; Rhodes, Gale; Welch, David B Mark; Zwerger, Monika

    2010-01-01

    Lamin B receptor (LBR) is an integral membrane protein of the interphase nuclear envelope (NE). The N-terminal end resides in the nucleoplasm, binding to lamin B and heterochromatin, with the interactions disrupted during mitosis. The C-terminal end resides within the inner nuclear membrane, retreating with the ER away from condensing chromosomes during mitotic NE breakdown. Some of these properties are interpretable in terms of our current structural knowledge of LBR, but many of the structural features remain unknown. LBR apparently has an evolutionary history which brought together at least two ancient conserved structural domains (i.e., Tudor and sterol reductase). This convergence may have occurred with the emergence of the chordates and echinoderms. It is not clear what survival values have maintained LBR structure during evolution. But it seems likely that roles in post-mitotic nuclear reformation, interphase NE growth and compartmentalization of nuclear architecture might have provided some evolutionary advantage to preservation of the LBR gene. PMID:21327105

  3. Laminate article

    DOEpatents

    Williams, Robert K.; Paranthaman, Mariappan; Chirayil, Thomas G.; Lee, Dominic F.; Goyal, Amit; Feenstra, Roeland

    2002-01-01

    A laminate article comprises a substrate and a biaxially textured (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer over the substrate, wherein 0laminate article can include a layer of YBCO over the (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer. A layer of CeO.sub.2 between the YBCO layer and the (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer can also be include. Further included can be a layer of YSZ between the CeO.sub.2 layer and the (RE.sub.x A.sub.(1-x)).sub.2 O.sub.2-(x/2) buffer layer. The substrate can be a biaxially textured metal, such as nickel. A method of forming the laminate article is also disclosed.

  4. Chromatin and lamin A determine two different mechanical response regimes of the cell nucleus.

    PubMed

    Stephens, Andrew D; Banigan, Edward J; Adam, Stephen A; Goldman, Robert D; Marko, John F

    2017-01-05

    The cell nucleus must continually resist and respond to inter- and intracellular mechanical forces to transduce mechanical signals and maintain proper genome organization and expression. Altered nuclear mechanics are associated with many human diseases, including heart disease, progeria, and cancer. Chromatin and nuclear envelope A-type lamin proteins are known to be key nuclear mechanical components perturbed in these diseases, but their distinct mechanical contributions are not known. Here, we directly establish the separate roles of chromatin and lamin A/C and show that they determine two distinct mechanical regimes via micromanipulation of single isolated nuclei. Chromatin governs response to small extensions (<3 μm), and eu-/heterochromatin levels modulate the stiffness. In contrast, lamin A/C levels control nuclear strain stiffening at large extensions. These results can be understood through simulations of a polymeric shell and crosslinked polymer interior. Our results provide a framework for understanding the differential effects of chromatin and lamin A/C in cell nuclear mechanics and their alterations in disease.

  5. Chromatin and lamin A determine two different mechanical response regimes of the cell nucleus

    PubMed Central

    Stephens, Andrew D.; Banigan, Edward J.; Adam, Stephen A.; Goldman, Robert D.; Marko, John F.

    2017-01-01

    The cell nucleus must continually resist and respond to intercellular and intracellular mechanical forces to transduce mechanical signals and maintain proper genome organization and expression. Altered nuclear mechanics is associated with many human diseases, including heart disease, progeria, and cancer. Chromatin and nuclear envelope A-type lamin proteins are known to be key nuclear mechanical components perturbed in these diseases, but their distinct mechanical contributions are not known. Here we directly establish the separate roles of chromatin and lamin A/C and show that they determine two distinct mechanical regimes via micromanipulation of single isolated nuclei. Chromatin governs response to small extensions (<3 μm), and euchromatin/heterochromatin levels modulate the stiffness. In contrast, lamin A/C levels control nuclear strain stiffening at large extensions. These results can be understood through simulations of a polymeric shell and cross-linked polymer interior. Our results provide a framework for understanding the differential effects of chromatin and lamin A/C in cell nuclear mechanics and their alterations in disease. PMID:28057760

  6. Mammalian telomeres and their partnership with lamins

    PubMed Central

    Burla, Romina; La Torre, Mattia; Saggio, Isabella

    2016-01-01

    ABSTRACT Chromosome ends are complex structures, which require a panel of factors for their elongation, replication, and protection. We describe here the mechanics of mammalian telomeres, dynamics and maintainance in relation to lamins. Multiple biochemical connections, including association of telomeres to the nuclear envelope and matrix, of telomeric proteins to lamins, and of lamin-associated proteins to chromosome ends, underline the interplay between lamins and telomeres. Paths toward senescence, such as defective telomere replication, altered heterochromatin organization, and impaired DNA repair, are common to lamins' and telomeres' dysfunction. The convergence of phenotypes can be interpreted through a model of dynamic, lamin-controlled functional platforms dedicated to the function of telomeres as fragile sites. The features of telomeropathies and laminopathies, and of animal models underline further overlapping aspects, including the alteration of stem cell compartments. We expect that future studies of basic biology and on aging will benefit from the analysis of this telomere-lamina interplay. PMID:27116558

  7. Lamins as mediators of oxidative stress

    SciTech Connect

    Sieprath, Tom; Darwiche, Rabih; De Vos, Winnok H.

    2012-05-18

    Highlights: Black-Right-Pointing-Pointer The nuclear lamina defines structural and functional properties of the cell nucleus. Black-Right-Pointing-Pointer Lamina dysfunction leads to a broad spectrum of laminopathies. Black-Right-Pointing-Pointer Recent data is reviewed connecting laminopathies to oxidative stress. Black-Right-Pointing-Pointer A framework is proposed to explain interactions between lamins and oxidative stress. -- Abstract: The nuclear lamina defines both structural and functional properties of the eukaryotic cell nucleus. Mutations in the LMNA gene, encoding A-type lamins, lead to a broad spectrum of diseases termed laminopathies. While different hypotheses have been postulated to explain disease development, there is still no unified view on the mechanistic basis of laminopathies. Recent observations indicate that laminopathies are often accompanied by altered levels of reactive oxygen species and a higher susceptibility to oxidative stress at the cellular level. In this review, we highlight the role of reactive oxygen species for cell function and disease development in the context of laminopathies and present a framework of non-exclusive mechanisms to explain the reciprocal interactions between a dysfunctional lamina and altered redox homeostasis.

  8. Interaction between the inner nuclear membrane lamin B receptor and the heterochromatic methyl binding protein, MeCP2

    SciTech Connect

    Guarda, Alessia; Bolognese, Fabrizio; Bonapace, Ian Marc; Badaracco, Gianfranco

    2009-07-01

    The nuclear membrane has an important role for the dynamic regulation of the genome, besides the well-established cytoskeletal function. The nuclear lamina is emerging as an important player in the organization of the position and functional state of interphase chromosomes. Epigenetic modifications such as DNA methylation and histone modifications are required for genome reprogramming during development, tissue-specific gene expression and global gene silencing. The Methyl-CpG binding protein MeCP2 binds methyl-CpG dinucleotides in the mammalian genome and functions as a transcriptional repressor in vivo by interacting with Sin3A, thereby recruiting histone deacetylases (HDAC). MeCP2 also mediates the formation of higher-order chromatin structures contributing to determine the architectural organization of the nucleus. In this paper, we show that MeCP2 interacts in vitro and in vivo with the inner nuclear membrane protein LBR and that the unstructured aminoacidic sequence linking the MBD and TRD domains of MeCP2 is responsible for this association. The formation of an LBR-MeCP2 protein complex might help providing a molecular explanation to the distribution of part of the heterochromatin at the nuclear periphery linked to inner membrane.

  9. Deletion of MLIP (muscle-enriched A-type lamin-interacting protein) leads to cardiac hyperactivation of Akt/mammalian target of rapamycin (mTOR) and impaired cardiac adaptation.

    PubMed

    Cattin, Marie-Elodie; Wang, Jessica; Weldrick, Jonathan J; Roeske, Cassandra L; Mak, Esther; Thorn, Stephanie L; DaSilva, Jean N; Wang, Yibin; Lusis, Aldon J; Burgon, Patrick G

    2015-10-30

    Aging and diseases generally result from tissue inability to maintain homeostasis through adaptation. The adult heart is particularly vulnerable to disequilibrium in homeostasis because its regenerative abilities are limited. Here, we report that MLIP (muscle enriched A-type lamin-interacting protein), a unique protein of unknown function, is required for proper cardiac adaptation. Mlip(-/-) mice exhibited normal cardiac function despite myocardial metabolic abnormalities and cardiac-specific overactivation of Akt/mTOR pathways. Cardiac-specific MLIP overexpression led to an inhibition of Akt/mTOR, providing evidence of a direct impact of MLIP on these key signaling pathways. Mlip(-/-) hearts showed an impaired capacity to adapt to stress (isoproterenol-induced hypertrophy), likely because of deregulated Akt/mTOR activity. Genome-wide association studies showed a genetic association between Mlip and early response to cardiac stress, supporting the role of MLIP in cardiac adaptation. Together, these results revealed that MLIP is required for normal myocardial adaptation to stress through integrated regulation of the Akt/mTOR pathways. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Deletion of MLIP (Muscle-enriched A-type Lamin-interacting Protein) Leads to Cardiac Hyperactivation of Akt/Mammalian Target of Rapamycin (mTOR) and Impaired Cardiac Adaptation*

    PubMed Central

    Cattin, Marie-Elodie; Wang, Jessica; Weldrick, Jonathan J.; Roeske, Cassandra L.; Mak, Esther; Thorn, Stephanie L.; DaSilva, Jean N.; Wang, Yibin; Lusis, Aldon J.; Burgon, Patrick G.

    2015-01-01

    Aging and diseases generally result from tissue inability to maintain homeostasis through adaptation. The adult heart is particularly vulnerable to disequilibrium in homeostasis because its regenerative abilities are limited. Here, we report that MLIP (muscle enriched A-type lamin-interacting protein), a unique protein of unknown function, is required for proper cardiac adaptation. Mlip−/− mice exhibited normal cardiac function despite myocardial metabolic abnormalities and cardiac-specific overactivation of Akt/mTOR pathways. Cardiac-specific MLIP overexpression led to an inhibition of Akt/mTOR, providing evidence of a direct impact of MLIP on these key signaling pathways. Mlip−/− hearts showed an impaired capacity to adapt to stress (isoproterenol-induced hypertrophy), likely because of deregulated Akt/mTOR activity. Genome-wide association studies showed a genetic association between Mlip and early response to cardiac stress, supporting the role of MLIP in cardiac adaptation. Together, these results revealed that MLIP is required for normal myocardial adaptation to stress through integrated regulation of the Akt/mTOR pathways. PMID:26359501

  11. Hybrid composite laminate structures

    NASA Technical Reports Server (NTRS)

    Chamis, C. C.; Lark, R. F. (Inventor)

    1977-01-01

    An invention which relates to laminate structures and specifically to essentially anisotropic fiber composite laminates is described. Metal foils are selectively disposed within the laminate to produce increased resistance to high velocity impact, fracture, surface erosion, and other stresses within the laminate.

  12. Lamins of the sea lamprey (Petromyzon marinus) and the evolution of the vertebrate lamin protein family.

    PubMed

    Schilf, Paul; Peter, Annette; Hurek, Thomas; Stick, Reimer

    2014-07-01

    Lamin proteins are found in all metazoans. Most non-vertebrate genomes including those of the closest relatives of vertebrates, the cephalochordates and tunicates, encode only a single lamin. In teleosts and tetrapods the number of lamin genes has quadrupled. They can be divided into four sub-types, lmnb1, lmnb2, LIII, and lmna, each characterized by particular features and functional differentiations. Little is known when during vertebrate evolution these features have emerged. Lampreys belong to the Agnatha, the sister group of the Gnathostomata. They split off first within the vertebrate lineage. Analysis of the sea lamprey (Petromyzon marinus) lamin complement presented here, identified three functional lamin genes, one encoding a lamin LIII, indicating that the characteristic gene structure of this subtype had been established prior to the agnathan/gnathostome split. Two other genes encode lamins for which orthology to gnathostome lamins cannot be designated. Search for lamin gene sequences in all vertebrate taxa for which sufficient sequence data are available reveals the evolutionary time frame in which specific features of the vertebrate lamins were established. Structural features characteristic for A-type lamins are not found in the lamprey genome. In contrast, lmna genes are present in all gnathostome lineages suggesting that this gene evolved with the emergence of the gnathostomes. The analysis of lamin gene neighborhoods reveals noticeable similarities between the different vertebrate lamin genes supporting the hypothesis that they emerged due to two rounds of whole genome duplication and makes clear that an orthologous relationship between a particular vertebrate paralog and lamins outside the vertebrate lineage cannot be established. Copyright © 2014 Elsevier GmbH. All rights reserved.

  13. Lamin A, farnesylation and aging

    SciTech Connect

    Reddy, Sita; Comai, Lucio

    2012-01-01

    Lamin A is a component of the nuclear envelope that is synthesized as a precursor prelamin A molecule and then processed into mature lamin A through sequential steps of posttranslational modifications and proteolytic cleavages. Remarkably, over 400 distinct point mutations have been so far identified throughout the LMNA gene, which result in the development of at least ten distinct human disorders, collectively known as laminopathies, among which is the premature aging disease Hutchinson-Gilford progeria syndrome (HGPS). The majority of HGPS cases are associated with a single point mutation in the LMNA gene that causes the production of a permanently farnesylated mutant lamin A protein termed progerin. The mechanism by which progerin leads to premature aging and the classical HGPS disease phenotype as well as the relationship between this disorder and the onset of analogous symptoms during the lifespan of a normal individual are not well understood. Yet, recent studies have provided critical insights on the cellular processes that are affected by accumulation of progerin and have suggested that cellular alterations in the lamin A processing pathway leading to the accumulation of farnesylated prelamin A intermediates may play a role in the aging process in the general population. In this review we provide a short background on lamin A and its maturation pathway and discuss the current knowledge of how progerin or alterations in the prelamin A processing pathway are thought to influence cell function and contribute to human aging.

  14. Lamin A, farnesylation and aging.

    PubMed

    Reddy, Sita; Comai, Lucio

    2012-01-01

    Lamin A is a component of the nuclear envelope that is synthesized as a precursor prelamin A molecule and then processed into mature lamin A through sequential steps of posttranslational modifications and proteolytic cleavages. Remarkably, over 400 distinct point mutations have been so far identified throughout the LMNA gene, which result in the development of at least ten distinct human disorders, collectively known as laminopathies, among which is the premature aging disease Hutchinson-Gilford progeria syndrome (HGPS). The majority of HGPS cases are associated with a single point mutation in the LMNA gene that causes the production of a permanently farnesylated mutant lamin A protein termed progerin. The mechanism by which progerin leads to premature aging and the classical HGPS disease phenotype as well as the relationship between this disorder and the onset of analogous symptoms during the lifespan of a normal individual are not well understood. Yet, recent studies have provided critical insights on the cellular processes that are affected by accumulation of progerin and have suggested that cellular alterations in the lamin A processing pathway leading to the accumulation of farnesylated prelamin A intermediates may play a role in the aging process in the general population. In this review we provide a short background on lamin A and its maturation pathway and discuss the current knowledge of how progerin or alterations in the prelamin A processing pathway are thought to influence cell function and contribute to human aging.

  15. Sustained accumulation of prelamin A and depletion of lamin A/C both cause oxidative stress and mitochondrial dysfunction but induce different cell fates.

    PubMed

    Sieprath, Tom; Corne, Tobias D J; Nooteboom, Marco; Grootaert, Charlotte; Rajkovic, Andreja; Buysschaert, Benjamin; Robijns, Joke; Broers, Jos L V; Ramaekers, Frans C S; Koopman, Werner J H; Willems, Peter H G M; De Vos, Winnok H

    2015-01-01

    The cell nucleus is structurally and functionally organized by lamins, intermediate filament proteins that form the nuclear lamina. Point mutations in genes that encode a specific subset of lamins, the A-type lamins, cause a spectrum of diseases termed laminopathies. Recent evidence points to a role for A-type lamins in intracellular redox homeostasis. To determine whether lamin A/C depletion and prelamin A accumulation differentially induce oxidative stress, we have performed a quantitative microscopy-based analysis of reactive oxygen species (ROS) levels and mitochondrial membrane potential (Δψm) in human fibroblasts subjected to sustained siRNA-mediated knockdown of LMNA and ZMPSTE24, respectively. We measured a highly significant increase in basal ROS levels and an even more prominent rise of induced ROS levels in lamin A/C depleted cells, eventually resulting in Δψm hyperpolarization and apoptosis. Depletion of ZMPSTE24 on the other hand, triggered a senescence pathway that was associated with moderately increased ROS levels and a transient Δψm depolarization. Both knockdowns were accompanied by an upregulation of several ROS detoxifying enzymes. Taken together, our data suggest that both persistent prelamin A accumulation and lamin A/C depletion elevate ROS levels, but to a different extent and with different effects on cell fate. This may contribute to the variety of disease phenotypes witnessed in laminopathies.

  16. Sustained accumulation of prelamin A and depletion of lamin A/C both cause oxidative stress and mitochondrial dysfunction but induce different cell fates

    PubMed Central

    Sieprath, Tom; Corne, Tobias DJ; Nooteboom, Marco; Grootaert, Charlotte; Rajkovic, Andreja; Buysschaert, Benjamin; Robijns, Joke; Broers, Jos LV; Ramaekers, Frans CS; Koopman, Werner JH; Willems, Peter HGM; De Vos, Winnok H

    2015-01-01

    The cell nucleus is structurally and functionally organized by lamins, intermediate filament proteins that form the nuclear lamina. Point mutations in genes that encode a specific subset of lamins, the A-type lamins, cause a spectrum of diseases termed laminopathies. Recent evidence points to a role for A-type lamins in intracellular redox homeostasis. To determine whether lamin A/C depletion and prelamin A accumulation differentially induce oxidative stress, we have performed a quantitative microscopy-based analysis of reactive oxygen species (ROS) levels and mitochondrial membrane potential (Δψm) in human fibroblasts subjected to sustained siRNA-mediated knockdown of LMNA and ZMPSTE24, respectively. We measured a highly significant increase in basal ROS levels and an even more prominent rise of induced ROS levels in lamin A/C depleted cells, eventually resulting in Δψm hyperpolarization and apoptosis. Depletion of ZMPSTE24 on the other hand, triggered a senescence pathway that was associated with moderately increased ROS levels and a transient Δψm depolarization. Both knockdowns were accompanied by an upregulation of several ROS detoxifying enzymes. Taken together, our data suggest that both persistent prelamin A accumulation and lamin A/C depletion elevate ROS levels, but to a different extent and with different effects on cell fate. This may contribute to the variety of disease phenotypes witnessed in laminopathies. PMID:25996284

  17. Farriery for chronic laminitis.

    PubMed

    O'Grady, Stephen E

    2010-08-01

    Laminitis is considered chronic once the distal phalanx has displaced within the hoof capsule. Chronic laminitis generally occurs as a direct sequel to acute laminitis. Clinical evaluation of chronic laminitis is best performed with a thorough clinical examination and radiography. The mainstay of hoof care is therapeutic farriery. In this article, the goals and principles of hoof care, the appropriate trim and various shoes that form the bulk of farriery for chronic laminitis, and surgical treatments are discussed.

  18. The tail domain of lamin B1 is more strongly modulated by divalent cations than lamin A

    PubMed Central

    Ganesh, Sairaam; Qin, Zhao; Spagnol, Stephen T; Biegler, Matthew T; Coffey, Kelli A; Kalinowski, Agnieszka; Buehler, Markus J; Dahl, Kris Noel

    2015-01-01

    The nucleoskeleton contains mainly nuclear intermediate filaments made of lamin proteins. Lamins provide nuclear structure and also play a role in various nuclear processes including signal transduction, transcription regulation and chromatin organization. The disparate functions of lamins may be related to the intrinsic disorder of the tail domains, which allows for altered and promiscuous binding. Here, we show modulation of lamin tail domain structures in the presence of divalent cations. We utilize changes in fluorescence of tryptophan residues within the Ig-fold flanked by disordered regions to experimentally measure protein thermodynamics. Using spectroscopy experiments and molecular dynamics simulations, we show that the tail domain of lamin B1 shows enhanced association with both Ca2+ and Mg2+ compared to the tail domain of lamin A. Binding curves show a similar KD between protein and ion (250–300 μM) for both proteins with both ions. However, we observe a maximum binding of ions to lamin B1 tail domain which is 2–3 times greater than that for lamin A tail domain by both experiment and simulation. Using simulations, we show that divalent ion association alters the Ig-fold by pinning flanking regions. With cells in culture, we observe altered lamin B1 organization in the presence of excess Mg2+ more so than for lamin A. We suggest that the differential sensitivity to divalent cations contributes to the vastly different functionalities and binding of the 2 proteins. PMID:25807068

  19. A Novel Lamin A Mutant Responsible for Congenital Muscular Dystrophy Causes Distinct Abnormalities of the Cell Nucleus.

    PubMed

    Barateau, Alice; Vadrot, Nathalie; Vicart, Patrick; Ferreiro, Ana; Mayer, Michèle; Héron, Delphine; Vigouroux, Corinne; Buendia, Brigitte

    2017-01-01

    A-type lamins, the intermediate filament proteins participating in nuclear structure and function, are encoded by LMNA. LMNA mutations can lead to laminopathies such as lipodystrophies, premature aging syndromes (progeria) and muscular dystrophies. Here, we identified a novel heterozygous LMNA p.R388P de novo mutation in a patient with a non-previously described severe phenotype comprising congenital muscular dystrophy (L-CMD) and lipodystrophy. In culture, the patient's skin fibroblasts entered prematurely into senescence, and some nuclei showed a lamina honeycomb pattern. C2C12 myoblasts were transfected with a construct carrying the patient's mutation; R388P-lamin A (LA) predominantly accumulated within the nucleoplasm and was depleted at the nuclear periphery, altering the anchorage of the inner nuclear membrane protein emerin and the nucleoplasmic protein LAP2-alpha. The mutant LA triggered a frequent and severe nuclear dysmorphy that occurred independently of prelamin A processing, as well as increased histone H3K9 acetylation. Nuclear dysmorphy was not significantly improved when transfected cells were treated with drugs disrupting microtubules or actin filaments or modifying the global histone acetylation pattern. Therefore, releasing any force exerted at the nuclear envelope by the cytoskeleton or chromatin did not rescue nuclear shape, in contrast to what was previously shown in Hutchinson-Gilford progeria due to other LMNA mutations. Our results point to the specific cytotoxic effect of the R388P-lamin A mutant, which is clinically related to a rare and severe multisystemic laminopathy phenotype.

  20. NMCP/LINC proteins: putative lamin analogs in plants?

    PubMed

    Ciska, Malgorzata; Moreno Diaz de la Espina, Susana

    2013-01-01

    Lamins are the main components of the metazoan lamina, and while the organization of the nuclear lamina of metazoans and plants is similar, there are apparently no genes encoding lamins or most lamin-binding proteins in plants. Thus, the plant lamina is not lamin-based and the proteins that form this structure are still to be characterized. Members of the plant NMCP/LINC/CRWN protein family share the typical tripartite structure of lamins, although the 2 exhibit no sequence similarity. However, given the many similarities between NMCP/LINC/CRWN proteins and lamins (structural organization, position of conserved regions, sub-nuclear distribution, solubility, and pattern of expression), these proteins are good candidates to carry out the functions of lamins in plants. Moreover, functional analysis of NMCP/LINC mutants has revealed their involvement in maintaining nuclear size and shape, another activity fulfilled by lamins. This review summarizes the current understanding of NMCP/LINC proteins and discusses future studies that will be required to demonstrate definitively that these proteins are plant analogs of lamins.

  1. Diverse lamin-dependent mechanisms interact to control chromatin dynamics

    PubMed Central

    Camozzi, Daria; Capanni, Cristina; Cenni, Vittoria; Mattioli, Elisabetta; Columbaro, Marta; Squarzoni, Stefano; Lattanzi, Giovanna

    2014-01-01

    Interconnected functional strategies govern chromatin dynamics in eukaryotic cells. In this context, A and B type lamins, the nuclear intermediate filaments, act on diverse platforms involved in tissue homeostasis. On the nuclear side, lamins elicit large scale or fine chromatin conformational changes, affect DNA damage response factors and transcription factor shuttling. On the cytoplasmic side, bridging-molecules, the LINC complex, associate with lamins to coordinate chromatin dynamics with cytoskeleton and extra-cellular signals.   Consistent with such a fine tuning, lamin mutations and/or defects in their expression or post-translational processing, as well as mutations in lamin partner genes, cause a heterogeneous group of diseases known as laminopathies. They include muscular dystrophies, cardiomyopathy, lipodystrophies, neuropathies, and progeroid syndromes. The study of chromatin dynamics under pathological conditions, which is summarized in this review, is shedding light on the complex and fascinating role of the nuclear lamina in chromatin regulation. PMID:25482195

  2. Direct actin binding to A- and B-type lamin tails and actin filament bundling by the lamin A tail

    PubMed Central

    Simon, Dan N; Zastrow, Michael S

    2010-01-01

    Nuclear intermediate filament networks formed by A- and B-type lamins are major components of the nucleoskeleton. Lamins have growing links to human physiology and disease including Emery-Dreifuss muscular dystrophy (EDMD), lipodystrophy, cardiomyopathy, neuropathy, cerebellar disorders and segmental accelerated ‘aging’ syndromes. How lamins interact with other nucleoskeletal components, and even the identities of these other components, are open questions. Previous studies suggested lamins might bind actin. We report that the recombinant C-terminal tail domain of human A- and B-type lamins binds directly to purified actin in high-speed pelleting assays. This interaction maps to a conserved Actin Binding site (AB-1) comprising lamin A residues 461–536 in the Ig-fold domain, which are 54% identical in lamin B1. Two EDMD-causing missense mutations (R527P and L530P) in lamin A that are predicted to disrupt the Ig-fold, each reduced F-actin binding by ∼66%, whereas the surface-exposed lipodystrophy-causing R482Q mutation had no significant effect. The lamin A tail was unique among lamins in having a second actin-binding site (AB-2). This second site was mapped to lamin A tail residues 564–608, based on actin-binding results for the lamin C tail and internal deletions in the lamin A tail that cause Hutchinson-Gilford Progeria Syndrome (Δ35, Δ50) or restrictive dermopathy (Δ90). Supporting the presence of two actin-binding sites, recombinant precursor (unmodified) and mature lamin A tails (not C or B1 tails) each bundled F-actin in vitro: furthermore F-actin bundling was reduced 25–40% by the R527P, L530P, Δ35 and Δ50 mutations, and was abolished by Δ90. Unexpectedly, the mature lamin A tail bound F-actin significantly more efficiently than did the prelamin A tail; this suggested unmodified residues 647–664, unique to prelamin A, might auto-inhibit binding to actin (and potentially other partners). These biochemical results suggest direct mechanisms

  3. Identification of differential protein interactors of lamin A and progerin

    PubMed Central

    Kubben, Nard; Voncken, Jan Willem; Demmers, Jeroen; Calis, Chantal; van Almen, Geert

    2010-01-01

    The nuclear lamina is an interconnected meshwork of intermediate filament proteins underlying the nuclear envelope. The lamina is an important regulator of nuclear structural integrity as well as nuclear processes, including transcription, DNA replication and chromatin remodeling. The major components of the lamina are A- and B-type lamins. Mutations in lamins impair lamina functions and cause a set of highly tissue-specific diseases collectively referred to as laminopathies. The phenotypic diversity amongst laminopathies is hypothesized to be caused by mutations affecting specific protein interactions, possibly in a tissue-specific manner. Current technologies to identify interaction partners of lamin A and its mutants are hampered by the insoluble nature of lamina components. To overcome the limitations of current technologies, we developed and applied a novel, unbiased approach to identify lamin A-interacting proteins. This approach involves expression of the high-affinity OneSTrEP-tag, precipitation of lamin-protein complexes after reversible protein cross-linking and subsequent protein identification by mass spectrometry. We used this approach to identify in mouse embryonic fibroblasts and cardiac myocyte NklTAg cell lines proteins that interact with lamin A and its mutant isoform progerin, which causes the premature aging disorder Hutchinson-Gilford progeria syndrome (HGPS). We identified a total of 313 lamina-interacting proteins, including several novel lamin A interactors, and we characterize a set of 35 proteins which preferentially interact with lamin A or progerin. PMID:21327095

  4. Lamination cooling system

    DOEpatents

    Rippel, Wally E.; Kobayashi, Daryl M.

    2005-10-11

    An electric motor, transformer or inductor having a lamination cooling system including a stack of laminations, each defining a plurality of apertures at least partially coincident with apertures of adjacent laminations. The apertures define a plurality of cooling-fluid passageways through the lamination stack, and gaps between the adjacent laminations are sealed to prevent a liquid cooling fluid in the passageways from escaping between the laminations. The gaps are sealed by injecting a heat-cured sealant into the passageways, expelling excess sealant, and heat-curing the lamination stack. The apertures of each lamination can be coincident with the same-sized apertures of adjacent laminations to form straight passageways, or they can vary in size, shape and/or position to form non-axial passageways, angled passageways, bidirectional passageways, and manifold sections of passageways that connect a plurality of different passageway sections. Manifold members adjoin opposite ends of the lamination stack, and each is configured with one or more cavities to act as a manifold to adjacent passageway ends. Complex manifold arrangements can create bidirectional flow in a variety of patterns.

  5. Distinct structural and mechanical properties of the nuclear lamina in Hutchinson-Gilford progeria syndrome.

    PubMed

    Dahl, Kris Noel; Scaffidi, Paola; Islam, Mohammad F; Yodh, Arjun G; Wilson, Katherine L; Misteli, Tom

    2006-07-05

    The nuclear lamina is a network of structural filaments, the A and B type lamins, located at the nuclear envelope and throughout the nucleus. Lamin filaments provide the nucleus with mechanical stability and support many basic activities, including gene regulation. Mutations in LMNA, the gene encoding A type lamins, cause numerous human diseases, including the segmental premature aging disease Hutchinson-Gilford progeria syndrome (HGPS). Here we show that structural and mechanical properties of the lamina are altered in HGPS cells. We demonstrate by live-cell imaging and biochemical analysis that lamins A and C become trapped at the nuclear periphery in HGPS patient cells. Using micropipette aspiration, we show that the lamina in HGPS cells has a significantly reduced ability to rearrange under mechanical stress. Based on polarization microscopy results, we suggest that the lamins are disordered in the healthy nuclei, whereas the lamins in HGPS nuclei form orientationally ordered microdomains. The reduced deformability of the HGPS nuclear lamina possibly could be due to the inability of these orientationally ordered microdomains to dissipate mechanical stress. Surprisingly, intact HGPS cells exhibited a degree of resistance to acute mechanical stress similar to that of cells from healthy individuals. Thus, in contrast to the nuclear fragility seen in lmna null cells, the lamina network in HGPS cells has unique mechanical properties that might contribute to disease phenotypes by affecting responses to mechanical force and misregulation of mechanosensitive gene expression.

  6. Distinct structural and mechanical properties of the nuclear lamina in Hutchinson–Gilford progeria syndrome

    PubMed Central

    Dahl, Kris Noel; Scaffidi, Paola; Islam, Mohammad F.; Yodh, Arjun G.; Wilson, Katherine L.; Misteli, Tom

    2006-01-01

    The nuclear lamina is a network of structural filaments, the A and B type lamins, located at the nuclear envelope and throughout the nucleus. Lamin filaments provide the nucleus with mechanical stability and support many basic activities, including gene regulation. Mutations in LMNA, the gene encoding A type lamins, cause numerous human diseases, including the segmental premature aging disease Hutchinson–Gilford progeria syndrome (HGPS). Here we show that structural and mechanical properties of the lamina are altered in HGPS cells. We demonstrate by live-cell imaging and biochemical analysis that lamins A and C become trapped at the nuclear periphery in HGPS patient cells. Using micropipette aspiration, we show that the lamina in HGPS cells has a significantly reduced ability to rearrange under mechanical stress. Based on polarization microscopy results, we suggest that the lamins are disordered in the healthy nuclei, whereas the lamins in HGPS nuclei form orientationally ordered microdomains. The reduced deformability of the HGPS nuclear lamina possibly could be due to the inability of these orientationally ordered microdomains to dissipate mechanical stress. Surprisingly, intact HGPS cells exhibited a degree of resistance to acute mechanical stress similar to that of cells from healthy individuals. Thus, in contrast to the nuclear fragility seen in lmna null cells, the lamina network in HGPS cells has unique mechanical properties that might contribute to disease phenotypes by affecting responses to mechanical force and misregulation of mechanosensitive gene expression. PMID:16801550

  7. Laminitis in the horse.

    PubMed

    Hood, D M

    1999-08-01

    This article serves as an introduction to this issue on laminitis. As such, it contains the general perspectives and terminology that will be used in all subsequent articles. This article separates the clinical problem of laminitis into developmental, acute, subacute, and chronic phases and defines the criteria, duration, clinical goals, and implications of these phases. The basis for the significance of laminitis to the horse industry and the horseman is reviewed. Lastly, the organization of this issue is described.

  8. A Novel Role of Lamins from Genetic Disease to Cancer Biomarkers

    PubMed Central

    Sakthivel, Kunnathur Murugesan; Sehgal, Poonam

    2016-01-01

    Lamins are the key components of the nuclear lamina and by virtue of their interactions with chromatin and binding partners act as regulators of cell proliferation and differentiation. Of late, the diverse roles of lamins in cellular processes have made them the topic of intense debate for their role in cancer progression. The observations about aberrant localization or misexpression of the nuclear lamins in cancerous tissues have often led to the speculative role of lamins as a cancer risk biomarker. Here we discuss the involvement of lamins in several cancer subtypes and their potential role in predicting the tumor progression. PMID:27994771

  9. A carboxyl-terminal interaction of lamin B1 is dependent on the CAAX endoprotease Rce1 and carboxymethylation.

    PubMed

    Maske, Christopher P; Hollinshead, Michael S; Higbee, Niall C; Bergo, Martin O; Young, Stephen G; Vaux, David J

    2003-09-29

    The mammalian nuclear lamina protein lamin B1 is posttranslationally modified by farnesylation, endoproteolysis, and carboxymethylation at a carboxyl-terminal CAAX motif. In this work, we demonstrate that the CAAX endoprotease Rce1 is required for lamin B1 endoproteolysis, demonstrate an independent pool of proteolyzed but nonmethylated lamin B1, as well as fully processed lamin B1, in interphase nuclei, and show a role for methylation in the organization of lamin B1 into domains of the nuclear lamina. Deficiency in the endoproteolysis or methylation of lamin B1 results in loss of integrity and deformity of the nuclear lamina. These data show that the organization of the nuclear envelope and lamina is dependent on a mechanism involving the methylation of lamin B1, and they identify a potential mechanism of laminopathy involving a B-type lamin.

  10. Analysis of operation of filters for post-accident decontamination of pressurized rooms of a nuclear power plants with a type VVER-440 reactor

    NASA Astrophysics Data System (ADS)

    Zaichik, L. I.; Zeigarnik, Yu. A.; Rotinov, A. G.; Sidorov, A. S.; Silina, N. N.; Chalyi, R. F.

    2007-05-01

    Operation of filters of postaccident decontamination of pressurized rooms of a nuclear power plant with a type-VVER-440 reactor is analyzed. The distribution of radioactive nuclides over filter stages, the time variation of the thermal state of filter, and the characteristic features of the processes of sorption in the section of fine cleaning are considered.

  11. Photovoltaic-Panel Laminator

    NASA Technical Reports Server (NTRS)

    Keenan, R.

    1985-01-01

    Two-piece unit heats and presses protective layers to form laminate. Rubber diaphragm between upper and lower vacuum chambers alternates between neutral position and one that presses against solar-cell array, supplying distributed force necessary to press layers of laminate together. Encapsulation helps to protect cells from environment and to ensure long panel life while allowing efficient generation of electricity from Sunlight.

  12. Photovoltaic-Panel Laminator

    NASA Technical Reports Server (NTRS)

    Keenan, R.

    1985-01-01

    Two-piece unit heats and presses protective layers to form laminate. Rubber diaphragm between upper and lower vacuum chambers alternates between neutral position and one that presses against solar-cell array, supplying distributed force necessary to press layers of laminate together. Encapsulation helps to protect cells from environment and to ensure long panel life while allowing efficient generation of electricity from Sunlight.

  13. The telomeric protein AKTIP interacts with A- and B-type lamins and is involved in regulation of cellular senescence

    PubMed Central

    Burla, Romina; Carcuro, Mariateresa; Torre, Mattia La; Fratini, Federica; Crescenzi, Marco; D'Apice, Maria Rosaria; Spitalieri, Paola; Raffa, Grazia Daniela; Astrologo, Letizia; Lattanzi, Giovanna; Cundari, Enrico; Raimondo, Domenico; Biroccio, Annamaria; Gatti, Maurizio

    2016-01-01

    AKTIP is a shelterin-interacting protein required for replication of telomeric DNA. Here, we show that AKTIP biochemically interacts with A- and B-type lamins and affects lamin A, but not lamin C or B, expression. In interphase cells, AKTIP localizes at the nuclear rim and in discrete regions of the nucleoplasm just like lamins. Double immunostaining revealed that AKTIP partially co-localizes with lamin B1 and lamin A/C in interphase cells, and that proper AKTIP localization requires functional lamin A. In mitotic cells, AKTIP is enriched at the spindle poles and at the midbody of late telophase cells similar to lamin B1. AKTIP-depleted cells show senescence-associated markers and recapitulate several aspects of the progeroid phenotype. Collectively, our results indicate that AKTIP is a new player in lamin-related processes, including those that govern nuclear architecture, telomere homeostasis and cellular senescence. PMID:27512140

  14. The telomeric protein AKTIP interacts with A- and B-type lamins and is involved in regulation of cellular senescence.

    PubMed

    Burla, Romina; Carcuro, Mariateresa; Torre, Mattia La; Fratini, Federica; Crescenzi, Marco; D'Apice, Maria Rosaria; Spitalieri, Paola; Raffa, Grazia Daniela; Astrologo, Letizia; Lattanzi, Giovanna; Cundari, Enrico; Raimondo, Domenico; Biroccio, Annamaria; Gatti, Maurizio; Saggio, Isabella

    2016-08-01

    AKTIP is a shelterin-interacting protein required for replication of telomeric DNA. Here, we show that AKTIP biochemically interacts with A- and B-type lamins and affects lamin A, but not lamin C or B, expression. In interphase cells, AKTIP localizes at the nuclear rim and in discrete regions of the nucleoplasm just like lamins. Double immunostaining revealed that AKTIP partially co-localizes with lamin B1 and lamin A/C in interphase cells, and that proper AKTIP localization requires functional lamin A. In mitotic cells, AKTIP is enriched at the spindle poles and at the midbody of late telophase cells similar to lamin B1. AKTIP-depleted cells show senescence-associated markers and recapitulate several aspects of the progeroid phenotype. Collectively, our results indicate that AKTIP is a new player in lamin-related processes, including those that govern nuclear architecture, telomere homeostasis and cellular senescence. © 2016 The Authors.

  15. Differentiation of C2C12 myoblasts expressing lamin A mutated at a site responsible for Emery-Dreifuss muscular dystrophy is improved by inhibition of the MEK-ERK pathway and stimulation of the PI3-kinase pathway

    SciTech Connect

    Favreau, Catherine; Delbarre, Erwan; Courvalin, Jean-Claude; Buendia, Brigitte

    2008-04-01

    Mutation R453W in A-type lamins, that are major nuclear envelope proteins, generates Emery-Dreifuss muscular dystrophy. We previously showed that mouse myoblasts expressing R453W-lamin A incompletely exit the cell cycle and differentiate into myocytes with a low level of multinucleation. Here we attempted to improve differentiation by treating these cells with a mixture of PD98059, an extracellular-regulated kinase (ERK) kinase (also known as mitogen-activated kinase, MEK) inhibitor, and insulin-like growth factor-II, an activator of phosphoinositide 3-kinase. We show that mouse myoblasts expressing R453W-lamin A were sensitive to the drug treatment as shown by (i) an increase in multinucleation, (ii) downregulation of proliferation markers (cyclin D1, hyperphosphorylated Rb), (iii) upregulation of myogenin, and (iv) sustained activation of p21 and cyclin D3. However, nuclear matrix anchorage of p21 and cyclin D3 in a complex with hypophosphorylated Rb that is critical to trigger cell cycle arrest and myogenin induction was deficient and incompletely restored by drug treatment. As the turn-over of R453W-lamin A at the nuclear envelope was greatly enhanced, we propose that R453W-lamin A impairs the capacity of the nuclear lamina to serve as scaffold for substrates of the MEK-ERK pathway and for MyoD-induced proteins that play a role in the differentiation process.

  16. ERK1/2 MAP kinases promote cell cycle entry by rapid, kinase-independent disruption of retinoblastoma-lamin A complexes.

    PubMed

    Rodríguez, Javier; Calvo, Fernando; González, José M; Casar, Berta; Andrés, Vicente; Crespo, Piero

    2010-11-29

    As orchestrators of essential cellular processes like proliferation, ERK1/2 mitogen-activated protein kinase signals impact on cell cycle regulation. A-type lamins are major constituents of the nuclear matrix that also control the cell cycle machinery by largely unknown mechanisms. In this paper, we disclose a functional liaison between ERK1/2 and lamin A whereby cell cycle progression is regulated. We demonstrate that lamin A serves as a mutually exclusive dock for ERK1/2 and the retinoblastoma (Rb) protein. Our results reveal that, immediately after their postactivation entrance in the nucleus, ERK1/2 dislodge Rb from its interaction with lamin A, thereby facilitating its rapid phosphorylation and consequently promoting E2F activation and cell cycle entry. Interestingly, these effects are independent of ERK1/2 kinase activity. We also show that cellular transformation and tumor cell proliferation are dependent on the balance between lamin A and nuclear ERK1/2 levels, which determines Rb accessibility for phosphorylation/inactivation.

  17. ERK1/2 MAP kinases promote cell cycle entry by rapid, kinase-independent disruption of retinoblastoma–lamin A complexes

    PubMed Central

    Rodríguez, Javier; Calvo, Fernando; José; González, M.; Casar, Berta; Andrés, Vicente

    2010-01-01

    As orchestrators of essential cellular processes like proliferation, ERK1/2 mitogen-activated protein kinase signals impact on cell cycle regulation. A-type lamins are major constituents of the nuclear matrix that also control the cell cycle machinery by largely unknown mechanisms. In this paper, we disclose a functional liaison between ERK1/2 and lamin A whereby cell cycle progression is regulated. We demonstrate that lamin A serves as a mutually exclusive dock for ERK1/2 and the retinoblastoma (Rb) protein. Our results reveal that, immediately after their postactivation entrance in the nucleus, ERK1/2 dislodge Rb from its interaction with lamin A, thereby facilitating its rapid phosphorylation and consequently promoting E2F activation and cell cycle entry. Interestingly, these effects are independent of ERK1/2 kinase activity. We also show that cellular transformation and tumor cell proliferation are dependent on the balance between lamin A and nuclear ERK1/2 levels, which determines Rb accessibility for phosphorylation/inactivation. PMID:21115804

  18. Defective lamin A-Rb signaling in Hutchinson-Gilford Progeria Syndrome and reversal by farnesyltransferase inhibition.

    PubMed

    Marji, Jackleen; O'Donoghue, Seán I; McClintock, Dayle; Satagopam, Venkata P; Schneider, Reinhard; Ratner, Desiree; Worman, Howard J; Gordon, Leslie B; Djabali, Karima

    2010-06-15

    Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare premature aging disorder caused by a de novo heterozygous point mutation G608G (GGC>GGT) within exon 11 of LMNA gene encoding A-type nuclear lamins. This mutation elicits an internal deletion of 50 amino acids in the carboxyl-terminus of prelamin A. The truncated protein, progerin, retains a farnesylated cysteine at its carboxyl terminus, a modification involved in HGPS pathogenesis. Inhibition of protein farnesylation has been shown to improve abnormal nuclear morphology and phenotype in cellular and animal models of HGPS. We analyzed global gene expression changes in fibroblasts from human subjects with HGPS and found that a lamin A-Rb signaling network is a major defective regulatory axis. Treatment of fibroblasts with a protein farnesyltransferase inhibitor reversed the gene expression defects. Our study identifies Rb as a key factor in HGPS pathogenesis and suggests that its modulation could ameliorate premature aging and possibly complications of physiological aging.

  19. Tying up loose ends: telomeres, genomic instability and lamins

    PubMed Central

    Eissenberg, Joel C.

    2016-01-01

    On casual inspection, the eukaryotic nucleus is a deceptively simple organelle. Far from being a bag of chromatin, the nucleus is, in some ways, a structural and functional extension of the chromosomes it contains. Recently, interest has intensified in how chromosome compartmentalization and dynamics affect nuclear function. Different studies uncovered functional interactions between chromosomes and the filamentous nuclear meshwork comprised of lamin proteins. Here, we summarize recent research suggesting that telomeres, the capping structures that protect chromosome ends, are stabilized by lamin-binding and that alterations in nuclear lamins lead to defects in telomere compartmentalization, homeostasis and function. Telomere dysfunction contributes to the genomic instability that characterizes aging-related diseases, and might be an important factor in the pathophysiology of lamin-related diseases. PMID:27010504

  20. Honeycomb-laminate composite structure

    NASA Technical Reports Server (NTRS)

    Gilwee, W. J., Jr.; Parker, J. A. (Inventor)

    1977-01-01

    A honeycomb-laminate composite structure was comprised of: (1) a cellular core of a polyquinoxaline foam in a honeycomb structure, and (2) a layer of a noncombustible fibrous material impregnated with a polyimide resin laminated on the cellular core. A process for producing the honeycomb-laminate composite structure and articles containing the honeycomb-laminate composite structure is described.

  1. Lamin B distribution and association with peripheral chromatin revealed by optical sectioning and electron microscopy tomography

    PubMed Central

    1993-01-01

    We have used a combination of immunogold staining, optical sectioning light microscopy, intermediate voltage electron microscopy, and EM tomography to examine the distribution of lamin B over the nuclear envelope of CHO cells. Apparent inconsistencies between previously published light and electron microscopy studies of nuclear lamin staining were resolved. At light microscopy resolution, an apparent open fibrillar network is visualized. Colocalization of lamin B and nuclear pores demonstrates that these apparent fibrils, separated by roughly 0.5 micron, are anti-correlated with the surface distribution of nuclear pores; pore clusters lie between or adjacent to regions of heavy lamin B staining. Examination at higher, EM resolution reveals that this apparent lamin B network does not correspond to an actual network of widely spaced, discrete bundles of lamin filaments. Rather it reflects a quantitative variation in lamin staining over a roughly 0.5-micron size scale, superimposed on a more continuous but still complex distribution of lamin filaments, spatially heterogeneous on a 0.1-0.2-micron size scale. Interestingly, lamin B staining at this higher resolution is highly correlated to the underlying chromatin distribution. Heavy concentrations of lamin B directly "cap" the surface of envelope associated, large-scale chromatin domains. PMID:8276889

  2. Aberrant localization of lamin B receptor (LBR) in cellular senescence in human cells

    SciTech Connect

    Arai, Rumi; En, Atsuki; Ukekawa, Ryo; Miki, Kensuke; Fujii, Michihiko; Ayusawa, Dai

    2016-05-13

    5-Bromodeoxyuridine (BrdU), a thymidine analogue, induces cellular senescence in mammalian cells. BrdU induces cellular senescence probably through the regulation of chromatin because BrdU destabilizes or disrupts nucleosome positioning and decondenses heterochromatin. Since heterochromatin is tethered to the nuclear periphery through the interaction with the nuclear envelope proteins, we examined the localization of the several nuclear envelope proteins such as lamins, lamin-interacting proteins, nuclear pore complex proteins, and nuclear transport proteins in senescent cells. We have shown here that lamin B receptor (LBR) showed a change in localization in both BrdU-induced and replicative senescent cells.

  3. Flexible thermal laminate

    NASA Technical Reports Server (NTRS)

    Dawn, F. S.; Sauers, D. G.

    1977-01-01

    Lightweight flexible laminate of interwoven conducting and insulating yarns, designed to provide localized controlled heating for propellant tanks on space vehicles, is useful for nonspace applications where weight, bulk, and flexibility are critical concerns.

  4. Historical perspectives on laminitis.

    PubMed

    Wagner, I P; Heymering, H

    1999-08-01

    This article attempts to provide a historical perspective regarding equine laminitis. It is designed to cover, as completely as possible, the historical record of, and the research advances made, in regards to acute and chronic laminitis. With respect to the historical record, the names given to this disease, the postulated etiologies, and the various treatment protocols are discussed. This article demonstrates the historical longevity of this disease and establishes a background for the current understanding of the disease's pathologic mechanisms and treatments.

  5. Role of Lamin B1 in Chromatin Instability

    PubMed Central

    Butin-Israeli, Veronika; Adam, Stephen A.; Jain, Nikhil; Otte, Gabriel L.; Neems, Daniel; Wiesmüller, Lisa; Berger, Shelly L.

    2014-01-01

    Nuclear lamins play important roles in the organization and structure of the nucleus; however, the specific mechanisms linking lamin structure to nuclear functions are poorly defined. We demonstrate that reducing nuclear lamin B1 expression by short hairpin RNA-mediated silencing in cancer cell lines to approximately 50% of normal levels causes a delay in the cell cycle and accumulation of cells in early S phase. The S phase delay appears to be due to the stalling and collapse of replication forks. The double-strand DNA breaks resulting from replication fork collapse were inefficiently repaired, causing persistent DNA damage signaling and the assembly of extensive repair foci on chromatin. The expression of multiple factors involved in DNA replication and repair by both nonhomologous end joining and homologous repair is misregulated when lamin B1 levels are reduced. We further demonstrate that lamin B1 interacts directly with the promoters of some genes associated with DNA damage response and repair, including BRCA1 and RAD51. Taken together, the results suggest that the maintenance of lamin B1 levels is required for DNA replication and repair through regulation of the expression of key factors involved in these essential nuclear functions. PMID:25535332

  6. Investigation of the entry characteristics of dust samplers of a type used in the British nuclear industry

    NASA Astrophysics Data System (ADS)

    Mark, D.; Vincent, J. H.; Stevens, D. C.; Marshall, M.

    Experiments have been carried out in a large wind tunnel to investigate the entry characteristics of dust samplers—both static and personal—of the type used to monitor 'total' airborne radioactive paniculate in the British nuclear industry. These samplers were exposed to test dusts of closely-graded fused alumina under experimental conditions relevant to the environmental conditions found in nuclear industry workplaces. For the static samplers (60-mm open face filter holders), performance was determined by reference to a 10-mm isokinetic probe. The resultant aspiration efficiency ( A) was found to be close to unity for the range of environmental conditions found in the nuclear industry workplace and for particles with aerodynamic diameter up to about 30 μm. Also it is unaffected by mounting the sampler itself on the large bluff body of the sampling pump. The performances of the personal samplers (of the 25-mm filter holder type) were assessed in terms of the ratio ( R) between the mass of dust entering each personal sampler when worn on the body of a mannequin and that entering the mouth of the mannequin under simulated breathing. The results show that, for nuclear industry workplace conditions, the personal samplers reflect satisfactorily the health-related 'total' dust exposure of the wearer.

  7. Chromosomal aneuploidies induced upon Lamin B2 depletion are mislocalized in the interphase nucleus.

    PubMed

    Ranade, Devika; Koul, Shivsmriti; Thompson, Joyce; Prasad, Kumar Brajesh; Sengupta, Kundan

    2017-03-01

    Chromosome territories assume non-random positions in the interphase nucleus with gene-rich chromosomes localized toward the nuclear interior and gene-poor chromosome territories toward the nuclear periphery. Lamins are intermediate filament proteins of the inner nuclear membrane required for the maintenance of nuclear structure and function. Here, we show using whole-genome expression profiling that Lamin A/C or Lamin B2 depletion in an otherwise diploid colorectal cancer cell line (DLD1) deregulates transcript levels from specific chromosomes. Further, three-dimensional fluorescence in situ hybridization (3D-FISH) analyses of a subset of these transcriptionally deregulated chromosome territories revealed that the diploid chromosome territories in Lamin-depleted cells largely maintain conserved positions in the interphase nucleus in a gene-density-dependent manner. In addition, chromosomal aneuploidies were induced in ~25 % of Lamin A/C or Lamin B2-depleted cells. Sub-populations of these aneuploid cells consistently showed a mislocalization of the gene-rich aneuploid chromosome 19 territory toward the nuclear periphery, while gene-poor aneuploid chromosome 18 territory was mislocalized toward the nuclear interior predominantly upon Lamin B2 than Lamin A/C depletion. In addition, a candidate gene locus ZNF570 (Chr.19q13.12) significantly overexpressed upon Lamin B2 depletion was remarkably repositioned away from the nuclear lamina. Taken together, our studies strongly implicate an overarching role for Lamin B2 in the maintenance of nuclear architecture since loss of Lamin B2 relieves the spatial positional constraints required for maintaining conserved localization of aneuploid chromosome territories in the interphase nucleus.

  8. Laminated rare earth structure and method of making

    DOEpatents

    Senor, David J [West Richland, WA; Johnson, Roger N [Richland, WA; Reid, Bruce D [Pasco, WA; Larson, Sandra [Richland, WA

    2002-07-30

    A laminated structure having two or more layers, wherein at least one layer is a metal substrate and at least one other layer is a coating comprising at least one rare earth element. For structures having more than two layers, the coating and metal substrate layers alternate. In one embodiment of the invention, the structure is a two-layer laminate having a rare earth coating electrospark deposited onto a metal substrate. In another embodiment of the invention, the structure is a three-layer laminate having the rare earth coating electrospark deposited onto a first metal substrate and the coating subsequently abonded to a second metal substrate. The bonding of the coating to the second metal substrate may be accomplished by hot pressing, hot rolling, high deformation rate processing, or combinations thereof. The laminated structure may be used in nuclear components where reactivity control or neutron absorption is desired and in non-nuclear applications such as magnetic and superconducting films.

  9. Label-free mass spectrometry exploits dozens of detected peptides to quantify lamins in wildtype and knockdown cells.

    PubMed

    Swift, Joe; Harada, Takamasa; Buxboim, Amnon; Shin, Jae-Won; Tang, Hsin-Yao; Speicher, David W; Discher, Dennis E

    2013-01-01

    Label-free quantitation and characterization of proteins by mass spectrometry (MS) is now feasible, especially for moderately expressed structural proteins such as lamins that typically yield dozens of tryptic peptides from tissue cells. Using standard cell culture samples, we describe general algorithms for quantitative analysis of peptides identified in liquid chromatography tandem mass spectrometry (LC-MS/MS). The algorithms were foundational to the discovery that the absolute stoichiometry of A-type to B-type lamins scales with tissue stiffness (Swift et al., Science 2013). Isoform dominance helps make sense of why mutations and changes with age of mechanosensitive lamin-A,C only affect "stiff" tissues such as heart, muscle, bone, or even fat, but not brain. A Peak Ratio Fingerprinting (PRF) algorithm is elaborated here through its application to lamin-A,C knockdown. After demonstrating the large dynamic range of PRF using calibrated mixtures of human and mouse lysates, we validate measurements of partial knockdown with standard cell biology analyses using quantitative immunofluorescence and immunoblotting. Optimal sets of MS-detected peptides as determined by PRF demonstrate that the strongest peptide signals are not necessarily the most reliable for quantitation. After lamin-A,C knockdown, PRF computes an invariant set of "housekeeping" proteins as part of a broader proteomic analysis that also shows the proteome of mesenchymal stem cells (MSCs) is more broadly perturbed than that of a human epithelial cancer line (A549s), with particular variation in nuclear and cytoskeletal proteins. These methods offer exciting prospects for basic and clinical studies of lamin-A,C as well as other MS-detectable proteins.

  10. Conserved lamin A protein expression in differentiated cells in the earthworm Eudrilus eugeniae.

    PubMed

    Kalidas, Ramamoorthy M; Raja, Subramanian Elaiya; Mydeen, Sheik Abdul Kader Nagoor Meeran; Samuel, Selvan Christyraj Johnson Retnaraj; Durairaj, Selvan Christyraj Jackson; Nino, Gopi D; Palanichelvam, Karuppaiah; Vaithi, Arumugaswami; Sudhakar, Sivasubramaniam

    2015-09-01

    Lamin A is an intermediate filament protein found in most of the differentiated vertebrate cells but absent in stem cells. It shapes the skeletal frame structure beneath the inner nuclear membrane of the cell nucleus. As there are few studies of the expression of lamin A in invertebrates, in the present work, we have analyzed the sequence, immunochemical conservation and expression pattern of lamin A protein in the earthworm Eudrilus eugeniae, a model organism for tissue regeneration. The expression of lamin A has been confirmed in E. eugeniae by immunoblot. Its localization in the nuclear membrane has been observed by immunohistochemistry using two different rabbit anti-sera raised against human lamin A peptides, which are located at the C-terminus of the lamin A protein. These two antibodies detected 70 kDa lamin A protein in mice and a single 65 kDa protein in the earthworm. The Oct-4 positive undifferentiated blastemal tissues of regenerating earthworm do not express lamin A, while the Oct-4 negative differentiated cells express lamin A. This pattern was also confirmed in the earthworm prostate gland. The present study is the first evidence for the immunochemical identification of lamin A and Oct-4 in the earthworm. Along with the partial sequence obtained from the earthworm genome, the present results suggest that lamin A protein and its expression pattern is conserved from the earthworm to humans.

  11. Lamins regulate cell trafficking and lineage maturation of adult human hematopoietic cells

    PubMed Central

    Shin, Jae-Won; Spinler, Kyle R.; Swift, Joe; Chasis, Joel A.; Mohandas, Narla; Discher, Dennis E.

    2013-01-01

    Hematopoietic stem and progenitor cells, as well as nucleated erythroblasts and megakaryocytes, reside preferentially in adult marrow microenvironments whereas other blood cells readily cross the endothelial barrier into the circulation. Because the nucleus is the largest organelle in blood cells, we hypothesized that (i) cell sorting across microporous barriers is regulated by nuclear deformability as controlled by lamin-A and -B, and (ii) lamin levels directly modulate hematopoietic programs. Mass spectrometry-calibrated intracellular flow cytometry indeed reveals a lamin expression map that partitions human blood lineages between marrow and circulating compartments (P = 0.00006). B-type lamins are highly variable and predominate only in CD34+ cells, but migration through micropores and nuclear flexibility in micropipette aspiration both appear limited by lamin-A:B stoichiometry across hematopoietic lineages. Differentiation is also modulated by overexpression or knockdown of lamins as well as retinoic acid addition, which regulates lamin-A transcription. In particular, erythroid differentiation is promoted by high lamin-A and low lamin-B1 expression whereas megakaryocytes of high ploidy are inhibited by lamin suppression. Lamins thus contribute to both trafficking and differentiation. PMID:24191023

  12. Lamination cooling system formation method

    DOEpatents

    Rippel, Wally E [Altadena, CA; Kobayashi, Daryl M [Monrovia, CA

    2012-06-19

    An electric motor, transformer or inductor having a cooling system. A stack of laminations have apertures at least partially coincident with apertures of adjacent laminations. The apertures define straight or angled cooling-fluid passageways through the lamination stack. Gaps between the adjacent laminations are sealed by injecting a heat-cured sealant into the passageways, expelling excess sealant, and heat-curing the lamination stack. Manifold members adjoin opposite ends of the lamination stack, and each is configured with one or more cavities to act as a manifold to adjacent passageway ends. Complex manifold arrangements can create bidirectional flow in a variety of patterns.

  13. Lamination cooling system formation method

    DOEpatents

    Rippel, Wally E [Altadena, CA; Kobayashi, Daryl M [Monrovia, CA

    2009-05-12

    An electric motor, transformer or inductor having a cooling system. A stack of laminations have apertures at least partially coincident with apertures of adjacent laminations. The apertures define straight or angled cooling-fluid passageways through the lamination stack. Gaps between the adjacent laminations are sealed by injecting a heat-cured sealant into the passageways, expelling excess sealant, and heat-curing the lamination stack. Manifold members adjoin opposite ends of the lamination stack, and each is configured with one or more cavities to act as a manifold to adjacent passageway ends. Complex manifold arrangements can create bidirectional flow in a variety of patterns.

  14. Lamin A and lamin-associated polypeptide 2 (LAP-2) in human skin in the process of aging.

    PubMed

    Golubtsova, N N; Filippov, F N; Gunin, A G

    2016-01-01

    At present time, relationships between lamins and processes leading to aging are established. Mutations of genes of lamins lead to diseases, one of them is progeria. This disease is caused by violation of splaysing of lamin A gene and accumulation the farnezylated prelamin A (progerin) in the nucleus. LAP-2 is an important factor which regulates and stabilizes the lamin A. However, roles of lamin A and LAP-2 in behavior of population of dermal fibroblasts in relation to age were not examined. The aim of this research was to study A type lamin and LAP-2 in human skin at different ages. Lamin A and LAP-2 were detected in sections of the skin by indirect immunohistochemistry. The number of fibroblasts containing lamin A was gradually decreased from 90,4 to 76,9 % from 20 weeks of gestation to 85 years old. There were 32 % of dermal fibroblasts with positive staining for LAP-2 at the period from 20 weeks of gestation to 20 years old. From 21 to 40 years, 37,8 % of fibroblasts containing lamin A were found in the dermis. In age interval 41-85 years, 49-51 % of dermal fibroblasts had a positive staining for LAP-2. Content of lamin A in the nuclei of fibroblasts was almost constant from 20 weeks of gestation to 85 years old. Expression of LAP-2 in the nuclei of fibroblasts was reduced from birth to 20 years old but increased from 21 years old. Number of fibroblasts and PCNA+ fibroblasts in dermis was diminished with age. The most significant decrease in the number of fibroblasts was observed from 20 weeks of gestation to 20 years old. Results allow to assume the participation of lamin A and LAP-2 in triggering age-dependent decrease in the number of fibroblasts in the dermis in humans.

  15. A Novel Lamin A Mutant Responsible for Congenital Muscular Dystrophy Causes Distinct Abnormalities of the Cell Nucleus

    PubMed Central

    Barateau, Alice; Vadrot, Nathalie; Vicart, Patrick; Ferreiro, Ana; Mayer, Michèle; Héron, Delphine; Vigouroux, Corinne

    2017-01-01

    A-type lamins, the intermediate filament proteins participating in nuclear structure and function, are encoded by LMNA. LMNA mutations can lead to laminopathies such as lipodystrophies, premature aging syndromes (progeria) and muscular dystrophies. Here, we identified a novel heterozygous LMNA p.R388P de novo mutation in a patient with a non-previously described severe phenotype comprising congenital muscular dystrophy (L-CMD) and lipodystrophy. In culture, the patient’s skin fibroblasts entered prematurely into senescence, and some nuclei showed a lamina honeycomb pattern. C2C12 myoblasts were transfected with a construct carrying the patient’s mutation; R388P-lamin A (LA) predominantly accumulated within the nucleoplasm and was depleted at the nuclear periphery, altering the anchorage of the inner nuclear membrane protein emerin and the nucleoplasmic protein LAP2-alpha. The mutant LA triggered a frequent and severe nuclear dysmorphy that occurred independently of prelamin A processing, as well as increased histone H3K9 acetylation. Nuclear dysmorphy was not significantly improved when transfected cells were treated with drugs disrupting microtubules or actin filaments or modifying the global histone acetylation pattern. Therefore, releasing any force exerted at the nuclear envelope by the cytoskeleton or chromatin did not rescue nuclear shape, in contrast to what was previously shown in Hutchinson-Gilford progeria due to other LMNA mutations. Our results point to the specific cytotoxic effect of the R388P-lamin A mutant, which is clinically related to a rare and severe multisystemic laminopathy phenotype. PMID:28125586

  16. Coupled actin-lamin biopolymer networks and protecting DNA

    NASA Astrophysics Data System (ADS)

    Zhang, Tao; Rocklin, D. Zeb; Mao, Xiaoming; Schwarz, J. M.

    The mechanical properties of cells are largely determined by networks of semiflexible biopolymers forming the cytoskeleton. Similarly, the mechanical properties of cell nuclei are also largely determined by networks of semiflexible biopolymers forming the nuclear cytoskeleton. In particular, a network of filamentous lamin sits just inside the inner nuclear membrane to presumably protect the heart of the cell nucleus--the DNA. It has been demonstrated over the past decade that the actin cytoskeletal biopolymer network and the lamin biopolymer network are coupled via a sequence of proteins bridging the outer and inner nuclear membranes, known as the LINC complex. We, therefore, probe the consequences of such a coupling in a model biopolymer network system via numerical simulations to understand the resulting deformations in the lamin network in response to perturbations in the actin cytoskeletal network. We find, for example, that the force transmission across the coupled system can depend sensitively on the concentration of LINC complexes. Such study could have implications for mechanical mechanisms of the regulation of transcription since DNA couples to lamin via lamin-binding domains so that deformations in the lamin network may result in deformations in the DNA.

  17. The Meiotic Nuclear Lamina Regulates Chromosome Dynamics and Promotes Efficient Homologous Recombination in the Mouse

    PubMed Central

    Schmitt, Johannes; Göb, Eva; Baar, Johannes; Ortega, Sagrario; Benavente, Ricardo; Alsheimer, Manfred

    2013-01-01

    The nuclear lamina is the structural scaffold of the nuclear envelope and is well known for its central role in nuclear organization and maintaining nuclear stability and shape. In the past, a number of severe human disorders have been identified to be associated with mutations in lamins. Extensive research on this topic has provided novel important clues about nuclear lamina function. These studies have contributed to the knowledge that the lamina constitutes a complex multifunctional platform combining both structural and regulatory functions. Here, we report that, in addition to the previously demonstrated significance for somatic cell differentiation and maintenance, the nuclear lamina is also an essential determinant for germ cell development. Both male and female mice lacking the short meiosis-specific A-type lamin C2 have a severely defective meiosis, which at least in the male results in infertility. Detailed analysis revealed that lamin C2 is required for telomere-driven dynamic repositioning of meiotic chromosomes. Loss of lamin C2 affects precise synapsis of the homologs and interferes with meiotic double-strand break repair. Taken together, our data explain how the nuclear lamina contributes to meiotic chromosome behaviour and accurate genome haploidization on a mechanistic level. PMID:23382700

  18. Laminate armor and related methods

    SciTech Connect

    Chu, Henry S; Lillo, Thomas M; Zagula, Thomas M

    2013-02-26

    Laminate armor and methods of manufacturing laminate armor. Specifically, laminate armor plates comprising a commercially pure titanium layer and a titanium alloy layer bonded to the commercially pure titanium outer layer are disclosed, wherein an average thickness of the titanium alloy inner layer is about four times an average thickness of the commercially pure titanium outer layer. In use, the titanium alloy layer is positioned facing an area to be protected. Additionally, roll-bonding methods for manufacturing laminate armor plates are disclosed.

  19. Mouse B-Type Lamins Are Required for Proper Organogenesis But Not by Embryonic Stem Cells

    PubMed Central

    Kim, Youngjo; Sharov, Alexei A.; McDole, Katie; Cheng, Melody; Hao, Haiping; Fan, Chen-Ming; Gaiano, Nicholas; Ko, Minoru S. H.; Zheng, Yixian

    2012-01-01

    B-type lamins, the major components of the nuclear lamina, are believed to be essential for cell proliferation and survival. We found that mouse embryonic stem cells (ESCs) do not need any lamins for self-renewal and pluripotency. Although genome-wide lamin-B binding profiles correlate with reduced gene expression, such binding is not directly required for gene silencing in ESCs or trophectoderm cells. However, B-type lamins are required for proper organogenesis. Defects in spindle orientation in neural progenitor cells and migration of neurons probably cause brain disorganizations found in lamin-B null mice. Thus, our studies not only disprove several prevailing views of lamin-Bs but also establish a foundation for redefining the function of the nuclear lamina in the context of tissue building and homeostasis. PMID:22116031

  20. Abnormal development of the cerebral cortex and cerebellum in the setting of lamin B2 deficiency

    PubMed Central

    Coffinier, Catherine; Chang, Sandy Y.; Nobumori, Chika; Tu, Yiping; Farber, Emily A.; Toth, Julia I.; Fong, Loren G.; Young, Stephen G.

    2010-01-01

    Nuclear lamins are components of the nuclear lamina, a structural scaffolding for the cell nucleus. Defects in lamins A and C cause an array of human diseases, including muscular dystrophy, lipodystrophy, and progeria, but no diseases have been linked to the loss of lamins B1 or B2. To explore the functional relevance of lamin B2, we generated lamin B2-deficient mice and found that they have severe brain abnormalities resembling lissencephaly, with abnormal layering of neurons in the cerebral cortex and cerebellum. This neuronal layering abnormality is due to defective neuronal migration, a process that is dependent on the organized movement of the nucleus within the cell. These studies establish an essential function for lamin B2 in neuronal migration and brain development. PMID:20145110

  1. Laminates and reinforced metals

    NASA Technical Reports Server (NTRS)

    Chamis, C. C.

    1980-01-01

    A selective review is presented of the state of the art of metallic laminates and fiber reinforced metals called metallic matrix laminates (MMLs). Design and analysis procedures that are used for, and typical structural components that have been made from MMLs are emphasized. Selected MMLs, constituent materials, typical material properties and fabrication procedures are briefly described, including hybrids and superhybrids. Advantages, disadvantages, and special considerations required during design, analysis, and fabrication of MMLs are examined. Tabular and graphical data are included to illustrate key aspects of MMLs. Appropriate references are cited to provide a selective bibliography of a rapidly expanding and very promising research and development field.

  2. A peculiar lamin in a peculiar mammal: Expression of lamin LIII in platypus (Ornithorhynchus anatinus).

    PubMed

    Peter, Annette; Khandekar, Shaunak; Deakin, Janine E; Stick, Reimer

    2015-11-01

    Platypus (Ornithorhynchus anatinus) holds a unique phylogenetic position at the base of the mammalian lineage due to an amalgamation of mammalian and sauropsid-like features. Here we describe the set of four lamin genes for platypus. Lamins are major components of the nuclear lamina, which constitutes a main component of the nucleoskeleton and is involved in a wide range of nuclear functions. Vertebrate evolution was accompanied by an increase in the number of lamin genes from a single gene in their closest relatives, the tunicates and cephalochordates, to four genes in the vertebrate lineage. Of the four genes the LIII gene is characterized by the presence of two alternatively spliced CaaX-encoding exons. In amphibians and fish LIII is the major lamin protein in oocytes and early embryos. The LIII gene is conserved throughout the vertebrate lineage, with the notable exception of marsupials and placental mammals, which have lost the LIII gene. Here we show that platypus has retained an LIII gene, albeit with a significantly altered structure and with a radically different expression pattern. The platypus LIII gene contains only a single CaaX-encoding exon and the head domain together with coil 1a and part of coil1b of the platypus LIII protein is replaced by a novel short non-helical N-terminus. It is expressed exclusively in the testis. These features resemble those of male germ cell-specific lamins in placental mammals, in particular those of lamin C2. Our data suggest (i) that the specific functions of LIII, which it fulfills in all other vertebrates, is no longer required in mammals and (ii) once it had been freed from these functions has undergone structural alterations and has adopted a new functionality in monotremes.

  3. Transparent polymeric laminates

    NASA Technical Reports Server (NTRS)

    Parker, J. A.; Fohlen, G. M.; Sawko, P. M.

    1973-01-01

    Laminate prepared from epoxy-boroxine and phenolphthalein polycarbonate has high mechanical strength at elevated temperature and is resistant to impact, fire, and high-energy thermal radiation. Polycarbonate is prepared by reaction of phenolphthalein with phosgene in presence of amine catalyst and immiscible organic solvent phase.

  4. Crystal structures of the coil 2B fragment and the globular tail domain of human lamin B1

    SciTech Connect

    Ruan, Jianbin; Xu, Chao; Bian, Chuanbing; Lam, Robert; Wang, Jia-Pey; Kania, Joanna; Min, Jinrong; Zang, Jianye

    2012-07-18

    We present here the crystal structures of human lamin B1 globular tail domain and coiled 2B domain, which adopt similar folds to Ig-like domain and coiled-coil domain of lamin A, respectively. Despite the overall similarity, we found an extra intermolecular disulfide bond in the lamin B1 coil 2B domain, which does not exist in lamin A/C. In addition, the structural analysis indicates that interactions at the lamin B1 homodimer interface are quite different from those of lamin A/C. Thus our research not only reveals the diversely formed homodimers among lamin family members, but also sheds light on understanding the important roles of lamin B1 in forming the nuclear lamina matrix.

  5. How do mutations in lamins A and C cause disease?

    PubMed Central

    Worman, Howard J.; Courvalin, Jean-Claude

    2004-01-01

    Mutations in lamins A and C, nuclear intermediate-filament proteins in nearly all somatic cells, cause a variety of diseases that primarily affect striated muscle, adipocytes, or peripheral nerves or cause features of premature aging. Two new studies use lamin A/C–deficient mice, which develop striated muscle disease, as a model to investigate pathogenic mechanisms. These reports provide evidence for a stepwise process in which mechanically stressed cells first develop chromatin and nuclear envelope damage and then develop secondary alterations in the transcriptional activation of genes in adaptive and protective pathways. PMID:14755330

  6. 78 FR 19007 - Certain Products Having Laminated Packaging, Laminated Packaging, and Components Thereof...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-28

    ... COMMISSION Certain Products Having Laminated Packaging, Laminated Packaging, and Components Thereof.... 1337, on behalf of Lamina Packaging Innovations LLC of Longview, Texas. An amended complaint was filed... importation of certain products having laminated packaging, laminated packaging, and components thereof...

  7. Chronic laminitis: foot management.

    PubMed

    Morrison, Scott

    2010-08-01

    Laminitis is a disease of the suspensory apparatus of the distal phalanx, which can advance to the chronic stage with varying degrees of structural failure. Because the disease may ultimately lead to mechanical failure of the digit, a foot management plan is required to effectively and mechanically treat these cases. Many laminitis cases can be successfully rehabilitated back to athletic soundness, light use, breeding, or pasture soundness, whereas others suffer from permanent instability and never enjoy an acceptable level of comfort. To understand how to minimize damage in the acute laminitic foot or rehabilitate the chronic laminitic foot, the veterinarian should have an understanding of the normal supporting structures of the digit, the biomechanical forces acting on the foot, and the structural failure that results when these otherwise normal forces act on a diseased, damaged foot.

  8. Purification of Lamins and Soluble Fragments of NETs.

    PubMed

    Makarov, Alexandr A; Rizzotto, Andrea; Meinke, Peter; Schirmer, Eric C

    2016-01-01

    Lamins and associated nuclear envelope transmembrane proteins (NETs) present unique problems for biochemical studies. Lamins form insoluble intermediate filament networks, associate with chromatin, and are also connected via specific NETs to the cytoskeleton, thus further complicating their isolation and purification from mammalian cells. Adding to this complexity, NETs at the inner nuclear membrane function in three distinct environments: (a) their nucleoplasmic domain(s) can bind lamins, chromatin, and transcriptional regulators; (b) they possess one or more integral transmembrane domains; and (c) their lumenal domain(s) function in the unique reducing environment of the nuclear envelope/ER lumen. This chapter describes strategic considerations and protocols to facilitate biochemical studies of lamins and NET proteins in vitro. Studying these proteins in vitro typically involves first expressing specific polypeptide fragments in bacteria and optimizing conditions to purify each fragment. We describe parameters for choosing specific fragments and designing purification strategies and provide detailed purification protocols. Biochemical studies can provide fundamental knowledge including binding strengths and the molecular consequences of disease-causing mutations that will be essential to understand nuclear envelope-genome interactions and nuclear envelope linked disease mechanisms. © 2016 Elsevier Inc. All rights reserved.

  9. The LMNA mutation p.Arg321Ter associated with dilated cardiomyopathy leads to reduced expression and a skewed ratio of lamin A and lamin C proteins

    SciTech Connect

    Al-Saaidi, Rasha; Rasmussen, Torsten B.; Palmfeldt, Johan; Nissen, Peter H.; Beqqali, Abdelaziz; Hansen, Jakob; Pinto, Yigal M.; Boesen, Thomas; Mogensen, Jens; Bross, Peter

    2013-11-15

    Dilated cardiomyopathy (DCM) is a disease of the heart muscle characterized by cardiac chamber enlargement and reduced systolic function of the left ventricle. Mutations in the LMNA gene represent the most frequent known genetic cause of DCM associated with disease of the conduction systems. The LMNA gene generates two major transcripts encoding the nuclear lamina major components lamin A and lamin C by alternative splicing. Both haploinsuffiency and dominant negative effects have been proposed as disease mechanism for premature termination codon (PTC) mutations in LMNA. These mechanisms however are still not clearly established. In this study, we used a representative LMNA nonsense mutation, p.Arg321Ter, to shed light on the molecular disease mechanisms. Cultured fibroblasts from three DCM patients carrying this mutation were analyzed. Quantitative reverse transcriptase PCR and sequencing of these PCR products indicated that transcripts from the mutant allele were degraded by the nonsense-mediated mRNA decay (NMD) mechanism. The fact that no truncated mutant protein was detectable in western blot (WB) analysis strengthens the notion that the mutant transcript is efficiently degraded. Furthermore, WB analysis showed that the expression of lamin C protein was reduced by the expected approximately 50%. Clearly decreased lamin A and lamin C levels were also observed by immunofluorescence microscopy analysis. However, results from both WB and nano-liquid chromatography/mass spectrometry demonstrated that the levels of lamin A protein were more reduced suggesting an effect on expression of lamin A from the wild type allele. PCR analysis of the ratio of lamin A to lamin C transcripts showed unchanged relative amounts of lamin A transcript suggesting that the effect on the wild type allele was operative at the protein level. Immunofluorescence microscopy analysis showed no abnormal nuclear morphology of patient fibroblast cells. Based on these data, we propose that

  10. Thermally stable laminating resins

    NASA Technical Reports Server (NTRS)

    Jones, R. J.; Vaughan, R. W.; Burns, E. A.

    1972-01-01

    Improved thermally stable laminating resins were developed based on the addition-type pyrolytic polymerization. Detailed monomer and polymer synthesis and characterization studies identified formulations which facilitate press molding processing and autoclave fabrication of glass and graphite fiber reinforced composites. A specific resin formulation, termed P10P was utilized to prepare a Courtaulds HMS reinforced simulated airfoil demonstration part by an autoclave molding process.

  11. Laminated piezoelectric transformer

    NASA Technical Reports Server (NTRS)

    Vazquez Carazo, Alfredo (Inventor)

    2006-01-01

    A laminated piezoelectric transformer is provided using the longitudinal vibration modes for step-up voltage conversion applications. The input portions are polarized to deform in a longitudinal plane and are bonded to an output portion. The deformation of the input portions is mechanically coupled to the output portion, which deforms in the same longitudinal direction relative to the input portion. The output portion is polarized in the thickness direction relative its electrodes, and piezoelectrically generates a stepped-up output voltage.

  12. Laminates for Ballistic Protection

    DTIC Science & Technology

    1975-02-01

    Textile Research &Inginearing Division 1S. NUM11ER OF PAGES N I ,aN4MotN.lInNOfDoe ISANSUITY CLASS. (of this eport0) 1 ~~,fICATIONf DOVINGRA3DING5 WS...Kevlar and glass laminates using an areal density of 11.6 kg/m and a maximn load of 82N (200 lb.). The Kevlar lamintes survived 1000 cycles but

  13. Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection.

    PubMed

    Wei, Wenqiang; Wang, Hongju; Li, Xiaoya; Fang, Na; Yang, Shili; Liu, Hongyan; Kang, Xiaonan; Sun, Xiulian; Ji, Shaoping

    2016-05-07

    At present, the details of lamina alterations after baculovirus infection remain elusive. In this study, a lamin gene in the Sf9 cell line of Spodoptera frugiperda was cloned. The open reading frame (orf) of the Sf9 lamin was 1860 bp and encoded a protein with a molecular weight of 70 kDa. A transfection assay with a red fluorescence protein (rfp)-lamin fusion protein indicated that Sf9 lamin was localized in the nuclear rim. Transmission electron microscopy observations indicated that Autographa californica multiple nucleopolyhedrovirus (AcMNPV) nucleocapsids may pass through the nuclear envelope. Immunofluorescence assay indicated that the lamina showed a ruffled staining pattern with the formation of invaginations in the Sf9 cells infected with AcMNPV, while it was evenly distributed at the nuclear periphery of mock-infected cells. Western blotting results indicated that the total amount of lamin in the baculovirus-infected Sf9 cells was significantly decreased compared with the mock-infected cells. These results imply that AcMNPV infection induces structural and biochemical rearrangements of lamina of Sf9 cells.

  14. Laminated Fresnel lenses

    SciTech Connect

    Jebens, R.W.

    1980-04-01

    A fabrication method for making plastic-on-glass laminated Fresnel lenses is discussed. These Fresnel lenses are for application in an RCA solar photovoltaic concentrator array now in the prototype stage of development. This laminated Fresnel lens fabrication method consists of making a Dow Corning J RTV silastic rubber mold of a master lens array. This mold is used to vacuum cast only the lens facets onto a low-iron tempered-glass substrate with an epoxy resin such as Hysol 0S 1000, a bisphenol-A resin with a flexibilizer that is anhydride cured. Cast acrylic Fresnel lens arrays commercialy available have potential cleaning and abrasion problems, have very large thermal expansion, and have dimensional uncertainties in their manufacture. The laminated lens is dimensionally stable with low thermal expansion, has good cleaning characteristics, and is very inexpensive in materials cost. The measured transmission of such a lens on low-iron glass is 80.4% compared with 85.1% for a cast acrylic lens, and the optical quality is good enough for application in the 100X to 200X concentration range. An approach to making large lens arrays (3 by 6 ft) on a commercial scale is explored.

  15. Solar cell module lamination process

    DOEpatents

    Carey, Paul G.; Thompson, Jesse B.; Aceves, Randy C.

    2002-01-01

    A solar cell module lamination process using fluoropolymers to provide protection from adverse environmental conditions and thus enable more extended use of solar cells, particularly in space applications. A laminate of fluoropolymer material provides a hermetically sealed solar cell module structure that is flexible and very durable. The laminate is virtually chemically inert, highly transmissive in the visible spectrum, dimensionally stable at temperatures up to about 200.degree. C. highly abrasion resistant, and exhibits very little ultra-violet degradation.

  16. Significance of 1B and 2B domains in modulating elastic properties of lamin A

    PubMed Central

    Bera, Manindra; Ainavarapu, Sri Rama Koti; Sengupta, Kaushik

    2016-01-01

    Nuclear lamins are type V intermediate filament proteins which form an elastic meshwork underlying the inner nuclear membrane. Lamins directly contribute to maintain the nuclear shape and elasticity. More than 400 mutations have been reported in lamin A that are involved in diseases known as laminopathies. These mutations are scattered mainly in the lamin rod domain along with some in its C-terminal domain. The contribution of the rod domain towards the elasticity of lamin A molecule was hitherto unknown. Here, we have elucidated the significance of the 1B and 2B domains of the rod in modulating the elastic behavior of lamin A by single-molecule force spectroscopy. In addition, we have also studied the network forming capacity of these domains and their corresponding viscoelastic behavior. We have shown that the 1B domain has the ability to form a lamin-like network and resists larger deformation. However at the single-molecular level, both the domains have comparable mechanical properties. The self-assembly of the 1B domain contributes to the elasticity of the lamin A network. PMID:27301336

  17. Laminitis in the geriatric horse.

    PubMed

    Hunt, Robert J

    2002-12-01

    There are few diseases that instill a comparable sense of doom in the mind of a treating veterinarian as laminitis. There is a feeling of cautious optimism when a horse with laminitis responds favorably to treatment. Although this optimism all too often proves false when treating laminitic patients, management of the patient afflicted with chronic laminitis can be rewarding. Through diligent and careful client communication and instruction, many geriatric patients with chronic laminitis can be maintained for years as comfortable companions, for light riding use, or as productive breeding animals.

  18. Mechanics of metal matrix laminates

    SciTech Connect

    Teply, J.L.

    1996-12-31

    The mechanics of fracture and fatigue crack propagation of ARALL Laminates and Discontinuously Reinforced SiC Aluminum (DRA) laminates is studied. In t these two, supposedly different, material systems the fracture and fatigue performance is closely related to their capability to delaminate. The delamination shape, length, rate of growth, effect on fatigue and fracture toughness are analyzed in both lamination systems. The similarities between these two lamination systems fatigue and fracture failure modes are determined and summarized. Design for compression is also mentioned.

  19. Age-related changes in lamin A/C expression in cardiomyocytes.

    PubMed

    Afilalo, Jonathan; Sebag, Igal A; Chalifour, Lorraine E; Rivas, Daniel; Akter, Rahima; Sharma, Kamal; Duque, Gustavo

    2007-09-01

    Lamin A and C (A/C) are type V intermediate filaments that form the nuclear lamina. Lamin A/C mutations lead to reduced expression of lamin A/C and diverse phenotypes such as familial cardiomyopathies and accelerated aging syndromes. Normal aging is associated with reduced expression of lamin A/C in osteoblasts and dermal fibroblasts but has never been assessed in cardiomyocytes. Our objective was to compare the expression of lamin A/C in cardiomyocytes of old (24 mo) versus young (4 mo) C57Bl/6J mice using a well-validated mouse model of aging. Lamin B1 was used as a control. Immunohistochemical and immunofluorescence analyses showed reduced expression of lamin A/C in cardiomyocyte nuclei of old mice (proportion of nuclei expressing lamin A/C, 9% vs. 62%, P < 0.001). Lamin A/C distribution was scattered peripherally and perinuclear in old mice, whereas it was homogeneous throughout the nuclei in young mice. Western blot analyses confirmed reduced expression of lamin A/C in nuclear extracts of old mice (ratio of lamin A/C to B1, 0.6 vs. 1.2, P < 0.01). Echocardiographic studies showed increased left ventricular wall thickness with preserved cavity size (concentric remodeling), increased left ventricular mass, and a slight reduction in fractional shortening in old mice. This is the first study to show that normal aging is associated with reduced expression and altered distribution of lamin A/C in nuclei of cardiomyocytes.

  20. A General Study of Hybrid Composite Laminates.

    DTIC Science & Technology

    1977-12-01

    appeared to have little effect on the overall properties of a laminate. Hybrid composite laminates obey classical laminate theory and can, in certain ply configurations, develop considerable free edge effect stresses. (Author)

  1. Nuclear matrix, nuclear envelope and premature aging syndromes in a translational research perspective.

    PubMed

    Cau, Pierre; Navarro, Claire; Harhouri, Karim; Roll, Patrice; Sigaudy, Sabine; Kaspi, Elise; Perrin, Sophie; De Sandre-Giovannoli, Annachiara; Lévy, Nicolas

    2014-05-01

    Lamin A-related progeroid syndromes are genetically determined, extremely rare and severe. In the past ten years, our knowledge and perspectives for these diseases has widely progressed, through the progressive dissection of their pathophysiological mechanisms leading to precocious and accelerated aging, from the genes mutations discovery until therapeutic trials in affected children. A-type lamins are major actors in several structural and functional activities at the nuclear periphery, as they are major components of the nuclear lamina. However, while this is usually poorly considered, they also play a key role within the rest of the nucleoplasm, whose defects are related to cell senescence. Although nuclear shape and nuclear envelope deformities are obvious and visible events, nuclear matrix disorganization and abnormal composition certainly represent the most important causes of cell defects with dramatic pathological consequences. Therefore, lamin-associated diseases should be better referred as laminopathies instead of envelopathies, this later being too restrictive, considering neither the key structural and functional roles of soluble lamins in the entire nucleoplasm, nor the nuclear matrix contribution to the pathophysiology of lamin-associated disorders and in particular in defective lamin A processing-associated aging diseases. Based on both our understanding of pathophysiological mechanisms and the biological and clinical consequences of progeria and related diseases, therapeutic trials have been conducted in patients and were terminated less than 10 years after the gene discovery, a quite fast issue for a genetic disease. Pharmacological drugs have been repurposed and used to decrease the toxicity of the accumulated, unprocessed and truncated prelaminA in progeria. To date, none of them may be considered as a cure for progeria and these clinical strategies were essentially designed toward reducing a subset of the most dramatic and morbid features

  2. Thermal conductivity of graphene laminate.

    PubMed

    Malekpour, H; Chang, K-H; Chen, J-C; Lu, C-Y; Nika, D L; Novoselov, K S; Balandin, A A

    2014-09-10

    We have investigated thermal conductivity of graphene laminate films deposited on polyethylene terephthalate substrates. Two types of graphene laminate were studied, as deposited and compressed, in order to determine the physical parameters affecting the heat conduction the most. The measurements were performed using the optothermal Raman technique and a set of suspended samples with the graphene laminate thickness from 9 to 44 μm. The thermal conductivity of graphene laminate was found to be in the range from 40 to 90 W/mK at room temperature. It was found unexpectedly that the average size and the alignment of graphene flakes are more important parameters defining the heat conduction than the mass density of the graphene laminate. The thermal conductivity scales up linearly with the average graphene flake size in both uncompressed and compressed laminates. The compressed laminates have higher thermal conductivity for the same average flake size owing to better flake alignment. Coating plastic materials with thin graphene laminate films that have up to 600× higher thermal conductivity than plastics may have important practical implications.

  3. Self-Healing Laminate System

    NASA Technical Reports Server (NTRS)

    Beiermann, Brett A. (Inventor); Keller, Michael W. (Inventor); White, Scott R. (Inventor); Sottos, Nancy R. (Inventor)

    2016-01-01

    A laminate material may include a first flexible layer, and a self-healing composite layer in contact with the first flexible layer. The composite layer includes an elastomer matrix, a plurality of first capsules including a polymerizer, and a corresponding activator for the polymerizer. The laminate material may self-heal when subjected to a puncture or a tear.

  4. The Development of Laminated Armor

    DTIC Science & Technology

    1948-09-27

    band saw, By using a piece of soft wood under the laminate while it was being cut, fraying of the edge was reduced to a minimum, especially in paneJs of...c. ’ LEGEND L LUMITE (SARAN) * ALL LAMINATE Rf RAYON (FQRTISAN) 20-30 OZ/FT 0-- Rc RAYON (CELANESE) ... 20-30...... X× GwLOss GLASS FLOSS Ir

  5. Effect of lamin A/C knockdown on osteoblast differentiation and function.

    PubMed

    Akter, Rahima; Rivas, Daniel; Geneau, Graziello; Drissi, Hicham; Duque, Gustavo

    2009-02-01

    Recent studies have associated mutations in lamin A/C, a component of the nuclear lamina, with premature aging and severe bone loss. In this study, we hypothesized that reduced expression of lamin A/C has a negative impact on osteoblastogenesis and bone formation in vitro. We inhibited lamin A/C using increasing doses of lamin A/C siRNA in normal human osteoblasts and differentiating mesenchymal stem cells (MSCs). Untreated cells and cells treated with vehicle but without the siRNA-oligo were used as control. The level of effectiveness of siRNA was determined by RT-PCR, Western blot, and immunofluorescence. Nuclear blebbing, a typical finding of lamin A/C inhibition, was quantified using propidium iodine staining, and its effect on cell survival was determined using MTS-formazan. Furthermore, alizarin red and alkaline phosphatase staining were correlated with osteocalcin secretion and levels of expression of osteocalcin, osterix, bone sialoprotein, and Runx2. Finally, the nuclear binding activity of Runx2, an essential transcription factor for osteoblast differentiation, was assessed using ELISA and EMSA. A successful inhibitory effect on the lamin A/C gene at doses of 400-800 nM oligo was obtained without affecting cell survival. Whereas osteoblast function was significantly affected by lamin A/C inhibition, siRNA-treated MSC showed a higher incidence of nuclear changes, lower osteoblast differentiation, and enhanced adipocyte differentiation. Finally, lamin A/C knockdown reduced Runx2 nuclear binding activity without affecting Runx2 expression. In summary, our results indicate that lamin A/C is a new factor needed for osteoblast differentiation that plays an important role in the cellular mechanisms of age-related bone loss.

  6. Circulation autoantibody against Lamin A/C in patients with Sjögren's syndrome

    PubMed Central

    Chen, Peng; Yang, Chunhe; Gan, Xianfeng; Hussain, Muhammad; Xun, Yiping; Tian, Yaping; Du, Hongwu

    2016-01-01

    Lamin A/C proteins are major components of nuclear laminae and were encoded by the LMNA gene. Recent studies have found that in addition to provides nuclear-membrane strength; it also regulates the gene expression. Lamin A/C has been confirmed as an autoantigen in RA, SLE and vasculitis. Anti-Lamin A/C antibodies also have been found by indirect immunofluorescence method. In this study, we used various research methods to confirm Lamin A/C is an autoantigen in Han Chinese patients with confirmed Sjögren's syndrome (SS). To further investigate the relationship between the autoimmune disease antigens, we compared the amino acid sequence of Lamin A/C epitope and several common antigens' antigenic determinant. As a result, we found that Lamin A/C has similar epitopes with U1RNP. It means that the potential relationship exist between Lamin A/C and U1RNP. Clinical data we collected also showed that anti-Lamin A/C and anti-U1RNP antibodies always appear in same serum sample. Therefore, we speculated that cross-reaction may take place between antigen and potential antigen, which have similar epitope. Then, by epitope spreading, the potential antigen can be a new autoantigen. Our study provided a new thinking for further research about the relationship between autoantigens and their development mechanism in autoimmune diseases. PMID:27835913

  7. In vitro posttranslational modification of lamin B cloned from a human T-cell line.

    PubMed Central

    Pollard, K M; Chan, E K; Grant, B J; Sullivan, K F; Tan, E M; Glass, C A

    1990-01-01

    Autoimmune diseases are characterized by spontaneously occurring autoantibodies which have proven to be useful reagents for the characterization of specific nuclear proteins. Using a monoclonal autoantibody (72B9) derived from a murine lupus strain, we have cloned a cDNA from the human T-cell line MOLT-4, which encodes nuclear lamin B. The identity of the encoded protein as lamin B was established by both biochemical and immunological criteria. Inspection of the deduced amino acid sequence of lamin B revealed the presence in coil 1B of the alpha-helical domain of a leucine heptad repeat region. Analysis of mRNA in HL60 and MOLT-4 cells, which express only lamin B, or HeLa cells, which express all three major lamins (A, B, and C), together with the comigration of in vitro-translated product with isolated HeLa cell lamin B by two-dimensional gel electrophoresis, suggests that a single lamin B is expressed in mammalian somatic cells. In vitro translation with the cDNA clone revealed an EDTA-sensitive posttranslational modification which resulted in an increase in the apparent molecular weight to that equivalent to the native in vivo-synthesized lamin B protein. This in vitro modification included incorporation of a product of mevalonolactone and required an intact carboxy terminus. Images PMID:2325650

  8. Lamin A reassembly at the end of mitosis is regulated by its SUMO-interacting motif.

    PubMed

    Moriuchi, Takanobu; Kuroda, Masaki; Kusumoto, Fumiya; Osumi, Takashi; Hirose, Fumiko

    2016-03-01

    Modification of proteins with small ubiquitin-related modifier (SUMO; SUMOylation) is involved in the regulation of various biological processes. Recent studies have demonstrated that noncovalent associations between SUMOylated proteins and co-operative proteins containing SUMO-interacting motifs (SIMs) are important for the spatiotemporal organization of many protein complexes. In this study, we demonstrate that interactions between lamin A, a major component of the nuclear lamina, and SUMO isoforms are dependent on one of the four SIMs (SIM3) resided in lamin A polypeptide in vitro. Live cell imaging and immunofluorescence staining showed that SIM3 is required for accumulation of lamin A on the chromosomes during telophase, and subsequent evaluation of a panel of deletion mutants determined that a 156-amino acid region spanning the carboxyl-terminal Ig-fold domain of lamin A is sufficient for this accumulation. Notably, mutation of SIM3 abrogated the dephosphorylation of mitosis-specific phosphorylation at Ser-22 of lamin A, which normally occurs during telophase, and the subsequent nuclear lamina reorganization. Furthermore, expression of a conjugation-defective SUMO2 mutant, which was previously shown to inhibit endogenous SUMOylation in a dominant-negative manner, also impaired the accumulation of wild type lamin A on telophase chromosomes. These findings suggest that interactions between SIM3 of lamin A and a putative SUMO2-modified protein plays an important role in the reorganization of the nuclear lamina at the end of mitosis.

  9. Identification of lamin B–regulated chromatin regions based on chromatin landscapes

    PubMed Central

    Zheng, Xiaobin; Kim, Youngjo; Zheng, Yixian

    2015-01-01

    Lamins, the major structural components of the nuclear lamina (NL) found beneath the nuclear envelope, are known to interact with most of the nuclear peripheral chromatin in metazoan cells. Although NL–chromatin associations correlate with a repressive chromatin state, the role of lamins in tethering chromatin to NL and how such tether influences gene expression have remained challenging to decipher. Studies suggest that NL proteins regulate chromatin in a context-dependent manner. Therefore understanding the context of chromatin states based on genomic features, including chromatin–NL interactions, is important to the study of lamins and other NL proteins. By modeling genome organization based on combinatorial patterns of chromatin association with lamin B1, core histone modification, and core and linker histone occupancy, we report six distinct large chromatin landscapes, referred to as histone lamin landscapes (HiLands)-red (R), -orange (O), -yellow (Y), -green (G), -blue (B), and -purple (P), in mouse embryonic stem cells (mESCs). This HiLands model demarcates the previously mapped lamin-associated chromatin domains (LADs) into two HiLands, HiLands-B and HiLands-P, which are similar to facultative and constitutive heterochromatins, respectively. Deletion of B-type lamins in mESCs caused a reduced interaction between regions of HiLands-B and NL as measured by emerin–chromatin interaction. Our findings reveal the importance of analyzing specific chromatin types when studying the function of NL proteins in chromatin tether and regulation. PMID:25995381

  10. Internal Stresses in Laminated Construction

    NASA Technical Reports Server (NTRS)

    Heim, A L; Knauss, A C; Seutter, Louis

    1923-01-01

    This report reviews the procedure employed in an investigation of the sources and influence of internal stresses in laminated construction, and discusses the influence of shrinkage and swelling stresses caused by atmospheric conditions upon the tensile strength across grain in laminated construction with special reference to airplane propellers. The investigation covered three sources of internal stress, namely, the combination of plain-sawed and quarter-sawed material in the same construction, the gluing together of laminations of different moisture contents, and the gluing together of laminations of different densities. Glued specimens and free specimens, made up under various manufacturing conditions, were subjected to various climatic changes inducing internal stresses and then were tested.

  11. Sudden bending of cracked laminates

    NASA Technical Reports Server (NTRS)

    Sih, G. C.; Chen, E. P.

    1980-01-01

    A dynamic approximate laminated plate theory is developed with emphasis placed on obtaining effective solution for the crack configuration where the 1/square root of r stress singularity and the condition of plane strain are preserved. The radial distance r is measured from the crack edge. The results obtained show that the crack moment intensity tends to decrease as the crack length to laminate plate thickness is increased. Hence, a laminated plate has the desirable feature of stabilizing a through crack as it increases its length at constant load. Also, the level of the average load intensity transmitted to a through crack can be reduced by making the inner layers to be stiffer than the outer layers. The present theory, although approximate, is useful for analyzing laminate failure to crack propagation under dynamic load conditions.

  12. Vacuum lamination of photovoltaic modules

    NASA Technical Reports Server (NTRS)

    Burger, D. R.

    1982-01-01

    Vacuum lamination of terrestrial photovoltaic modules is a new high volume process requiring new equipment and newly develop materials. Equipment development, materials research, and some research in related fields and testing methods are discussed.

  13. Laminated electromagnetic pump stator core

    DOEpatents

    Fanning, Alan W.

    1995-01-01

    A stator core for an electromagnetic pump includes a plurality of circumferentially abutting tapered laminations extending radially outwardly from a centerline axis to collectively define a radially inner bore and a radially outer circumference. Each of the laminations includes radially inner and outer edges and has a thickness increasing from the inner edge toward the outer edge to provide a substantially continuous path adjacent the circumference.

  14. Metamaterial properties of periodic laminates

    NASA Astrophysics Data System (ADS)

    Srivastava, Ankit

    2016-11-01

    In this paper we show that a 1-D phononic crystal (laminate) can exhibit metamaterial wave phenomena which are traditionally associated with 2- and 3-D crystals. Moreover, due to the absence of a length scale in 2 of its dimensions, it can outperform higher dimensional crystals on some measures. This includes allowing only negative refraction over large frequency ranges and serving as a near-omnidirectional high-pass filter up to a large frequency value. First we provide a theoretical discussion on the salient characteristics of the dispersion relation of a laminate and formulate the solution of an interface problem by the application of the normal mode decomposition technique. We present a methodology with which to induce a pure negative refraction in the laminate. As a corollary to our approach of negative refraction, we show how the laminate can be used to steer beams over large angles for small changes in the incident angles (beam steering). Furthermore, we clarify how the transmitted modes in the laminate can be switched on and off by varying the angle of the incident wave by a small amount. Finally, we show that the laminate can be used as a remarkably efficient high-pass frequency filter. An appropriately designed laminate will reflect all plane waves from quasi-static to a large frequency, incident at it from all angles except for a small set of near-normal incidences. This will be true even if the homogeneous medium is impedance matched with the laminate. Due to the similarities between SH waves and electromagnetic (EM) waves it is expected that some or all of these results may also apply to EM waves in a layered periodic dielectric.

  15. Push Tester For Laminated Films

    NASA Technical Reports Server (NTRS)

    Sugimura, Russell S.

    1991-01-01

    Small instrument used to measure brittleness of polymer film adhesively bonded to hard substrate. Penlike instrument has microball tip. Small pointer in slot on side of instrument used to calibrate and indicate spring force applied by point. Microball dents only small area of specimen. Such measurements used to measure rates of embrittlement in environmental tests of candidate laminated-film covers for photovoltaic modules. Not limited to transparent films; also used on opaque laminated films on back panels of photovoltaic modules.

  16. Comparing lamin proteins post-translational relative stability using a 2A peptide-based system reveals elevated resistance of progerin to cellular degradation

    PubMed Central

    Wu, Di; Zhang, Haoyue; Cao, Kan

    2016-01-01

    ABSTRACT Nuclear lamins are the major components of the nuclear lamina at the periphery of the nucleus, supporting the nuclear envelope and participating in many nuclear processes, including DNA replication, transcription and chromatin organization. A group of diseases, the laminopathies, is associated with mutations in lamin genes. One of the most striking cases is Hutchinson-Gilford progeria syndrome (HGPS) which is the consequence of a lamin A dominant negative mutant named progerin. Due to the abnormal presence of a permanent C-terminal farnesyl tail, progerin gradually accumulates on the nuclear membrane, perturbing a diversity of signalings and transcriptional events. The accumulation of progerin has led to the speculation that progerin possesses higher stability than the wild type lamin A protein. However, the low solubility of lamin proteins renders traditional immunoprecipitation-dependent methods such as pulse-chase analysis ineffective for comparing the relative stabilities of mutant and wild type lamins. Here, we employ a novel platform for inferring differences in lamin stability, which is based on normalization to a co-translated reporter protein following porcine teschovirus-1 2A peptide-mediated co-translational cleavage. The results obtained using this method support the notion that progerin is more stable than lamin A. Moreover, treatment of FTI reduces progerin relative stability to the level of wild type lamin A. PMID:27929926

  17. Increased expression of the Hutchinson-Gilford progeria syndrome truncated lamin A transcript during cell aging.

    PubMed

    Rodriguez, Sofia; Coppedè, Fabio; Sagelius, Hanna; Eriksson, Maria

    2009-07-01

    Most cases of the segmental progeroid syndrome, Hutchinson-Gilford progeria syndrome (HGPS), are caused by a de novo dominant mutation within a single codon of the LMNA gene. This mutation leads to the increased usage of an internal splice site that generates an alternative lamin A transcript with an internal deletion of 150 nucleotides, called lamin A Delta 150. The LMNA gene encodes two major proteins of the inner nuclear lamina, lamins A and C, but not much is known about their expression levels. Determination of the overall expression levels of the LMNA gene transcripts is an important step to further the understanding of the HGPS. In this study, we have performed absolute quantification of the lamins A, C and A Delta 150 transcripts in primary dermal fibroblasts from HGPS patients and unaffected age-matched and parent controls. We show that the lamin A Delta 150 transcript is present in unaffected controls but its expression is >160-fold lower than that in samples from HGPS patients. Analysis of transcript expression during in vitro aging shows that although the levels of lamin A and lamin C transcripts remain unchanged, the lamin A Delta 150 transcript increases in late passage cells from HGPS patients and parental controls. This study provides a new method for LMNA transcript analysis and insights into the expression of the LMNA gene in HGPS and normal cells.

  18. Defective Lamin A-Rb Signaling in Hutchinson-Gilford Progeria Syndrome and Reversal by Farnesyltransferase Inhibition

    PubMed Central

    Marji, Jackleen; O'Donoghue, Seán I.; McClintock, Dayle; Satagopam, Venkata P.; Schneider, Reinhard; Ratner, Desiree; J. Worman, Howard; Gordon, Leslie B.; Djabali, Karima

    2010-01-01

    Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare premature aging disorder caused by a de novo heterozygous point mutation G608G (GGC>GGT) within exon 11 of LMNA gene encoding A-type nuclear lamins. This mutation elicits an internal deletion of 50 amino acids in the carboxyl-terminus of prelamin A. The truncated protein, progerin, retains a farnesylated cysteine at its carboxyl terminus, a modification involved in HGPS pathogenesis. Inhibition of protein farnesylation has been shown to improve abnormal nuclear morphology and phenotype in cellular and animal models of HGPS. We analyzed global gene expression changes in fibroblasts from human subjects with HGPS and found that a lamin A-Rb signaling network is a major defective regulatory axis. Treatment of fibroblasts with a protein farnesyltransferase inhibitor reversed the gene expression defects. Our study identifies Rb as a key factor in HGPS pathogenesis and suggests that its modulation could ameliorate premature aging and possibly complications of physiological aging. PMID:20559568

  19. Targeting Mitogen-Activated Protein Kinase Signaling in Mouse Models of Cardiomyopathy Caused by Lamin A/C Gene Mutations

    PubMed Central

    Muchir, Antoine; Worman, Howard J.

    2016-01-01

    The most frequently occurring mutations in the gene encoding nuclear lamin A and nuclear lamin C cause striated muscle diseases virtually always involving the heart. In this review, we describe the approaches and methods used to discover that cardiomyopathy-causing lamin A/C gene mutations increase MAP kinase signaling in the heart and that this plays a role in disease pathogenesis. We review different mouse models of cardiomyopathy caused by lamin A/C gene mutations and how transcriptomic analysis of one model identified increased cardiac activity of the ERK1/2, JNK, and p38α MAP kinases. We describe methods used to measure the activity of these MAP kinases in mouse hearts and then discuss preclinical treatment protocols using pharmacological inhibitors to demonstrate their role in pathogenesis. Several of these kinase inhibitors are in clinical development and could potentially be used to treat human subjects with cardiomyopathy caused by lamin A/C gene mutations. PMID:26795484

  20. Emerin organizes actin flow for nuclear movement and centrosome orientation in migrating fibroblasts.

    PubMed

    Chang, Wakam; Folker, Eric S; Worman, Howard J; Gundersen, Gregg G

    2013-12-01

    In migrating fibroblasts, rearward movement of the nucleus orients the centrosome toward the leading edge. Nuclear movement results from coupling rearward-moving, dorsal actin cables to the nucleus by linear arrays of nesprin-2G and SUN2, termed transmembrane actin-associated nuclear (TAN) lines. A-type lamins anchor TAN lines, prompting us to test whether emerin, a nuclear membrane protein that interacts with lamins and TAN line proteins, contributes to nuclear movement. In fibroblasts depleted of emerin, nuclei moved nondirectionally or completely failed to move. Consistent with these nuclear movement defects, dorsal actin cable flow was nondirectional in cells lacking emerin. TAN lines formed normally in cells lacking emerin and were coordinated with the erratic nuclear movements, although in 20% of the cases, TAN lines slipped over immobile nuclei. Myosin II drives actin flow, and depletion of myosin IIB, but not myosin IIA, showed similar nondirectional nuclear movement and actin flow as in emerin-depleted cells. Myosin IIB specifically coimmunoprecipitated with emerin, and emerin depletion prevented myosin IIB localization near nuclei. These results show that emerin functions with myosin IIB to polarize actin flow and nuclear movement in fibroblasts, suggesting a novel function for the nuclear envelope in organizing directional actin flow and cytoplasmic polarity.

  1. Bovine acidosis: implications on laminitis.

    PubMed

    Nocek, J E

    1997-05-01

    Bovine lactic acidosis syndrome is associated with large increases of lactic acid in the rumen, which result from diets that are high in ruminally available carbohydrates, or forage that is low in effective fiber, or both. The syndrome involves two separate anatomical areas, the gastrointestinal tract and body fluids, and is related to the rate and extent of lactic acid production, utilization, and absorption. Clinical manifestations range from loss of appetite to death. Lactic acid accumulates in the rumen when the bacteria that synthesize lactic acid outnumber those that utilize lactic acid. The systemic impact of acidosis may have several physiological implications, including laminitis, a diffuse aseptic inflammation of the laminae (corium). Although a nutritional basis for the disease exists, etiology includes a multitude of interactive factors, such as metabolic and digestive disorders, postpartum stress, and localized trauma, which lead to the release of vasoactive substances that trigger mechanisms that cause degenerative changes in the foot. The severity of laminitis is related to the frequency, intensity, and duration of systemic acidotic insults on the mechanisms responsible for the release of vasoactive substance. The critical link between acidosis and laminitis appears to be associated with a persistent hypoperfusion, which results in ischemia in the digit. Management of acidosis is critical in preventing laminitis. High producing dairy herds attempting to maximize energy intake are continually confronted with subclinical acidosis and laminitis. Management of feeding and husbandry practices can be implemented to reduce incidence of disease.

  2. Polymer metal lamination by radiation method

    NASA Astrophysics Data System (ADS)

    Yizheng, Ma; Maoqing, Chen; Zhixiong, Feng; Dakuan, Sun; Dawei, You

    1993-07-01

    Conventional technology of PE-metal laminate involves the use of adhesives for laminating. Environ pollution has been a big problem not easily resolved and processing is complicated. By radiation peroxidation of the surface of polyethylene a simple polyethylene-metal laminating method has been succeeded. Measurements of the mechanical properties and the electrical properties made according to China's national standards showed the properties of the laminating film conforming well with the national standards. It has been found that the adhesion strength of the PE-metal film is affected by irradiation, the metal surface reactivity and cleaning of the laminating surface. PE-metal film laminated by radiation method is more heat resistant than the film laminated by conventional methods. The laminate materials can be widely used in electrical devices, chemical and packaging industry.

  3. Dynamics of Lamin-A Processing Following Precursor Accumulation

    PubMed Central

    Liu, Qian; Kim, Dae In; Syme, Janet; LuValle, Phyllis; Burke, Brian; Roux, Kyle J.

    2010-01-01

    Lamin A (LaA) is a component of the nuclear lamina, an intermediate filament meshwork that underlies the inner nuclear membrane (INM) of the nuclear envelope (NE). Newly synthesized prelamin A (PreA) undergoes extensive processing involving C-terminal farnesylation followed by proteolysis yielding non-farnesylated mature lamin A. Different inhibitors of these processing events are currently used therapeutically. Hutchinson-Gilford Progeria Syndrome (HGPS) is most commonly caused by mutations leading to an accumulation of a farnesylated LaA isoform, prompting a clinical trial using farnesyltransferase inhibitors (FTI) to reduce this modification. At therapeutic levels, HIV protease inhibitors (PI) can unexpectedly inhibit the final processing step in PreA maturation. We have examined the dynamics of LaA processing and associated cellular effects during PI or FTI treatment and following inhibitor washout. While PI reversibility was rapid, with respect to both LaA maturation and associated cellular phenotype, recovery from FTI treatment was more gradual. FTI reversibility is influenced by both cell type and rate of proliferation. These results suggest a less static lamin network than has previously been observed. PMID:20526372

  4. Basic mechanics of laminated composite plates

    NASA Technical Reports Server (NTRS)

    Nettles, Alan T.

    1994-01-01

    The mechanics of laminated composite materials is presented in a clear manner with only essential derivations included. The constitutive equations in all of their forms are developed and then summarized in a separate section. The effects of hygrothermal effects are included. The prediction of the engineering constants for a laminate are derived. Strength of laminated composites is not covered.

  5. Strength and stiffness of small glued-laminated beams with different qualities of tension laminations

    Treesearch

    Catherine M. Marx; Russell C. Moody

    1981-01-01

    A total of 180 small Douglas Fir–Larch (DF-L) or Southern Pine (SP) glued-laminated beams were evaluated to determine the tension lamination quality necessary to obtain desired design stresses. The test beams had either the regular laminating grades of L1 DF-L/No. 1D SP or the special 302-24 laminating grade as tension laminations. Because an initial set of SP beams...

  6. A mechanism of AP-1 suppression through interaction of c-Fos with lamin A/C

    PubMed Central

    Ivorra, Carmen; Kubicek, Markus; González, José M.; Sanz-González, Silvia M.; Álvarez-Barrientos, Alberto; O'Connor, José-Enrique; Burke, Brian; Andrés, Vicente

    2006-01-01

    AP-1 (Activating Protein 1) transcription factor activity is tightly regulated at multiple levels, including dimer formation (i.e., Fos/Jun). Here we show that the intermediate filament protein lamin A/C suppresses AP-1 function through direct interaction with c-Fos, and that both proteins can interact and colocalize at the nuclear envelope (NE) in mammalian cells. Perinuclear localization of c-Fos is absent in Lmna-null cells but can be restored by lamin A overexpression. In vitro, preincubation of c-Fos with lamin A prior to the addition of c-Jun inhibits AP-1 DNA-binding activity. In vivo, overexpression of lamin A reduces the formation of c-Fos/c-Jun heterodimers, and suppresses AP-1 DNA-binding and transcriptional activity. Notably, c-Fos colocalizes with lamin A/C at the NE in starvation-synchronized quiescent cells lacking detectable AP-1 DNA binding. In contrast, serum-induced AP-1 DNA-binding activity coincides with abundant nucleoplasmic c-Fos expression without changes in lamin A/C localization. We also found that Lmna-null cells display enhanced proliferation. In contrast, lamin A overexpression causes growth arrest, and ectopic c-Fos partially overcomes lamin A/C-induced cell cycle alterations. We propose lamin A/C-mediated c-Fos sequestration at the NE as a novel mechanism of transcriptional and cell cycle control. PMID:16452503

  7. Lamin B1 protein is required for dendrite development in primary mouse cortical neurons.

    PubMed

    Giacomini, Caterina; Mahajani, Sameehan; Ruffilli, Roberta; Marotta, Roberto; Gasparini, Laura

    2016-01-01

    Lamin B1, a key component of the nuclear lamina, plays an important role in brain development and function. A duplication of the human lamin B1 (LMNB1) gene has been linked to adult-onset autosomal dominant leukodystrophy, and mouse and human loss-of-function mutations in lamin B1 are susceptibility factors for neural tube defects. In the mouse, experimental ablation of endogenous lamin B1 (Lmnb1) severely impairs embryonic corticogenesis. Here we report that in primary mouse cortical neurons, LMNB1 overexpression reduces axonal outgrowth, whereas deficiency of endogenous Lmnb1 results in aberrant dendritic development. In the absence of Lmnb1, both the length and complexity of dendrites are reduced, and their growth is unresponsive to KCl stimulation. This defective dendritic outgrowth stems from impaired ERK signaling. In Lmnb1-null neurons, ERK is correctly phosphorylated, but phospho-ERK fails to translocate to the nucleus, possibly due to delocalization of nuclear pore complexes (NPCs) at the nuclear envelope. Taken together, these data highlight a previously unrecognized role of lamin B1 in dendrite development of mouse cortical neurons through regulation of nuclear shuttling of specific signaling molecules and NPC distribution.

  8. Lamin B1 protein is required for dendrite development in primary mouse cortical neurons

    PubMed Central

    Giacomini, Caterina; Mahajani, Sameehan; Ruffilli, Roberta; Marotta, Roberto; Gasparini, Laura

    2016-01-01

    Lamin B1, a key component of the nuclear lamina, plays an important role in brain development and function. A duplication of the human lamin B1 (LMNB1) gene has been linked to adult-onset autosomal dominant leukodystrophy, and mouse and human loss-of-function mutations in lamin B1 are susceptibility factors for neural tube defects. In the mouse, experimental ablation of endogenous lamin B1 (Lmnb1) severely impairs embryonic corticogenesis. Here we report that in primary mouse cortical neurons, LMNB1 overexpression reduces axonal outgrowth, whereas deficiency of endogenous Lmnb1 results in aberrant dendritic development. In the absence of Lmnb1, both the length and complexity of dendrites are reduced, and their growth is unresponsive to KCl stimulation. This defective dendritic outgrowth stems from impaired ERK signaling. In Lmnb1-null neurons, ERK is correctly phosphorylated, but phospho-ERK fails to translocate to the nucleus, possibly due to delocalization of nuclear pore complexes (NPCs) at the nuclear envelope. Taken together, these data highlight a previously unrecognized role of lamin B1 in dendrite development of mouse cortical neurons through regulation of nuclear shuttling of specific signaling molecules and NPC distribution. PMID:26510501

  9. Chronic laminitis: current treatment strategies.

    PubMed

    Parks, Andrew; O'Grady, Stephen E

    2003-08-01

    Laminitis is divided into four different phases: developmental, acute, subacute, and chronic. The focus of this article is on treating the laminitic horse after the cessation of therapy for the acute phase, that is, usually 2 to 4 weeks after the onset of clinical signs.

  10. Laminitis as a systemic disease.

    PubMed

    Hood, D M

    1999-08-01

    This article presents the clinical pathology and the involvement of the cardiovascular, renal, endocrine, and immunologic systems in laminitis. The data available on these systems are presented with respect to the disease phase and severity. The nutritional and metabolic alterations realized in the chronically affected horse are also presented. In this discussion, the origins and clinical implications of these systemic findings are discussed.

  11. Multilayer printed wiring board lamination

    SciTech Connect

    Lula, J.W.

    1980-06-01

    The relationship of delamination resistance of multilayer PWBs made from GF material to manufacturing process variables was investigated. A unique quantitative test method developed during this project shows that delamination resistance is highly sensitive to material conditioning, to innerlayer surface treatment, and to post-lamination storage conditions, but is relatively insensitive to cure cycle variations.

  12. Steady state response of unsymmetrically laminated plates

    SciTech Connect

    Hosokawa, Kenji; Kawashima, Katsuya; Sakata, Toshiyuki

    1995-11-01

    A numerical approach for analyzing the forced vibration problem of a symmetrically laminated FRP (fiber reinforced plastic) composite plate was proposed by the authors. In the present paper, this approach is modified for application to an unsymmetrically laminated FRP composite plate. Numerical calculations are carried out for the clamped antisymmetrically laminated rectangular and elliptical plates which are a kind of unsymmetrically laminated plate. Then,, the effects of the lamina material and the fiber orientation angle on the steady state response are discussed. Furthermore, it is investigated that what structural damping factor is most influenced on the steady state response of an antisymmetrically laminated plate.

  13. The nucleoporin ELYS/Mel28 regulates nuclear envelope subdomain formation in HeLa cells

    PubMed Central

    Clever, Michaela; Funakoshi, Tomoko; Mimura, Yasuhiro; Takagi, Masatoshi; Imamoto, Naoko

    2012-01-01

    In open mitosis the nuclear envelope (NE) reassembles at the end of each mitosis. This process involves the reformation of the nuclear pore complex (NPC), the inner and outer nuclear membranes, and the nuclear lamina. In human cells cell cycle-dependent NE subdomains exist, characterized as A-type lamin-rich/NPC-free or B-type lamin-rich/NPC-rich, which are initially formed as core or noncore regions on mitotic chromosomes, respectively. Although postmitotic NE formation has been extensively studied, little is known about the coordination of NPC and NE assembly. Here, we report that the nucleoporin ELYS/Mel28, which is crucial for postmitotic NPC formation, is essential for recruiting the lamin B receptor (LBR) to the chromosomal noncore region. Furthermore, ELYS/Mel28 is responsible for focusing of A-type lamin-binding proteins like emerin, Lap2α and the barrier-to-autointegration factor (BAF) at the chromosomal core region. ELYS/Mel28 biochemically interacts with the LBR in a phosphorylation-dependent manner. Recruitment of the LBR depends on the nucleoporin Nup107, which interacts with ELYS/Mel28 but not on nucleoporin Pom121, suggesting that the specific molecular interactions with ELYS/Mel28 are involved in the NE assembly at the noncore region. The depletion of the LBR affected neither the behavior of emerin nor Lap2α indicating that the recruitment of the LBR to mitotic chromosomes is not involved in formation of the core region. The depletion of ELYS/Mel28 also accelerates the entry into cytokinesis after recruitment of emerin to chromosomes. Our data show that ELYS/Mel28 plays a role in NE subdomain formation in late mitosis. PMID:22555603

  14. Structure and stability of the lamin A tail domain and HGPS mutant.

    PubMed

    Qin, Zhao; Kalinowski, Agnieszka; Dahl, Kris Noel; Buehler, Markus J

    2011-09-01

    Hutchinson-Gilford progeria syndrome (HGPS) is a premature aging syndrome caused by the expression and accumulation of a mutant form of lamin A, Δ50 lamin A. As a component of the cell's nucleoskeleton, lamin A plays an important role in the mechanical stabilization of the nuclear envelope and in other nuclear functions. It is largely unknown how the characteristic 50 amino acid deletion affects the conformation of the mostly intrinsically disordered tail domain of lamin A. Here we perform replica exchange molecular dynamics simulations of the tail domain and determine an ensemble of semi-stable structures. Based on these structures we show that the ZMPSTE 24 cleavage site on the precursor form of the lamin A tail domain orients itself in such a way as to facilitate cleavage during the maturation process. We confirm our simulated structures by comparing the thermodynamic properties of the ensemble structures to in vitro stability measurements. Using this combination of experimental and computational techniques, we compare the size, heterogeneity of size, thermodynamic stability of the Ig-fold, as well as the mechanisms of force-induced denaturation. Our data shows that the Δ50 lamin A tail domain is more compact and displays less heterogeneity than the mature lamin A tail domain. Altogether these results suggest that the altered structure and stability of the tail domain can explain changed protein-protein and protein-DNA interactions and may represent an etiology of the disease. Also, this study provides the first molecular structure(s) of the lamin A tail domain, which is confirmed by thermodynamic tests in experiment.

  15. Lamination residual strains and stresses in hybrid laminates

    NASA Technical Reports Server (NTRS)

    Daniel, I. M.; Liber, T.

    1977-01-01

    An investigation is conducted of the effects of hybridization on the magnitude of lamination residual stresses. Eight-ply graphite/Kevlar 49/epoxy and graphite/S-glass/epoxy laminates were studied. The same matrix resin was selected for all basic materials to ensure compatibility and uniform curing of the various plies. The specimens, with inserted strain gages and thermocouples, were subjected to curing and postcuring cycles in an autoclave. Subsequently, the specimens were subjected to a thermal cycle from room temperature to 444 K and down to room temperature. It was found that hydridizing reduces apparently residual strains and stresses in the graphite plies. However, these strains were not affected much by the type and degree of hybridization.

  16. Lamin B1 is required for mature neuron-specific gene expression during olfactory sensory neuron differentiation

    PubMed Central

    Gigante, Crystal M.; Dibattista, Michele; Dong, Frederick N.; Zheng, Xiaobin; Yue, Sibiao; Young, Stephen G.; Reisert, Johannes; Zheng, Yixian; Zhao, Haiqing

    2017-01-01

    B-type lamins are major constituents of the nuclear lamina in all metazoan cells, yet have specific roles in the development of certain cell types. Although they are speculated to regulate gene expression in developmental contexts, a direct link between B-type lamins and developmental gene expression in an in vivo system is currently lacking. Here, we identify lamin B1 as a key regulator of gene expression required for the formation of functional olfactory sensory neurons. By using targeted knockout in olfactory epithelial stem cells in adult mice, we show that lamin B1 deficient neurons exhibit attenuated response to odour stimulation. This deficit can be explained by decreased expression of genes involved in mature neuron function, along with increased expression of genes atypical of the olfactory lineage. These results support that the broadly expressed lamin B1 regulates expression of a subset of genes involved in the differentiation of a specific cell type. PMID:28425486

  17. What Should the Cardiologist know about Lamin Disease?

    PubMed Central

    Charron, Philippe; Arbustini, Eloisa; Bonne, Gisèle

    2012-01-01

    Lamins are intermediate filament proteins able to polymerise and form an organised meshwork underlying the inner nuclear membrane in most differentiated somatic cells. Mutations in the LMNA gene, which encodes the two major lamin A and C isoforms, cause a diverse range of diseases, called laminopathies, including dilated cardiomyopathy, associated with a poor prognosis and high rate of sudden death due to conduction defect and early ventricular arrhythmia. Identification of mutations in LMNA gene in clinical practice is rapidly increasing, as well as comprehensive cardiac and genetic family screening. As a consequence, cardiologists are more and more frequently faced to difficult questions regarding optimal management of patients and relatives, especially timing for prophylactic cardioverter defibrillator. This review focuses on recent data useful for the clinician, as well as therapeutic perspectives both in human and animal models. PMID:26835025

  18. A computational procedure to analyze metal matrix laminates with nonlinear lamination residual strains

    NASA Technical Reports Server (NTRS)

    Chamis, C. C.; Sullivan, T. L.

    1974-01-01

    An approximate computational procedure is described for the analysis of angleplied laminates with residual nonlinear strains. The procedure consists of a combination of linear composite mechanics and incremental linear laminate theory. The procedure accounts for initial nonlinear strains, unloading, and in-situ matrix orthotropic nonlinear behavior. The results obtained in applying the procedure to boron/aluminum angleplied laminates show that this is a convenient means to accurately predict the initial tangent properties of angleplied laminates in which the matrix has been strained nonlinearly by the lamination residual stresses. The procedure predicted initial tangent properties results which were in good agreement with measured data obtained from boron/aluminum angleplied laminates.

  19. Mechanics in human fibroblasts and progeria: Lamin A mutation E145K results in stiffening of nuclei.

    PubMed

    Apte, Ketaki; Stick, Reimer; Radmacher, Manfred

    2017-02-01

    The lamina is a filamentous meshwork beneath the inner nuclear membrane that confers mechanical stability to nuclei. The E145K mutation in lamin A causes Hutchinson-Gilford progeria syndrome (HGPS). It affects lamin filament assembly and induces profound changes in the nuclear architecture. Expression of wild-type and E145K lamin A in Xenopus oocytes followed by atomic force microscopy (AFM) probing of isolated oocyte nuclei has shown significant changes in the mechanical properties of the lamina. Nuclei of oocytes expressing E145K lamin A are stiffer than those expressing wild-type lamin A. Here we present mechanical measurements by AFM on dermal fibroblasts obtained from a 4-year-old progeria patient bearing the E145K lamin A mutation and compared it to fibroblasts obtained from 2 healthy donors of 10 and 61 years of age, respectively. The abnormal shape of nuclei expressing E145K lamin A was analyzed by fluorescence microscopy. Lamina thickness was measured using electron micrographs. Fluorescence microscopy showed alterations in the actin network of progeria cells. AFM probing of whole dermal fibroblasts did not demonstrate significant differences in the elastic moduli of nuclear and cytoplasmic cell regions. In contrast, AFM measurements of isolated nuclei showed that nuclei of progeria and old person's cells are significantly stiffer than those of the young person, indicating that the process of aging, be it natural or abnormal, increases nuclear stiffness. Our results corroborate AFM data obtained using Xenopus oocyte nuclei and prove that the presence of E145K lamin A abnormally increases nuclear stiffness.

  20. Plastic Laminate Pulsed Power Development

    SciTech Connect

    ALEXANDER,JEFF A.; SHOPE,STEVEN L.; PATE,RONALD C.; RINEHART,LARRY F.; JOJOLA,JOHN M.; RUEBUSH,MITCHELL H.; CROWE,WAYNE; LUNDSTROM,J.; SMITH,T.; ZAGAR,D.; PRESTWICH,K.

    2000-09-01

    The desire to move high-energy Pulsed Power systems from the laboratory to practical field systems requires the development of compact lightweight drivers. This paper concerns an effort to develop such a system based on a plastic laminate strip Blumlein as the final pulseshaping stage for a 600 kV, 50ns, 5-ohm driver. A lifetime and breakdown study conducted with small-area samples identified Kapton sheet impregnated with Propylene Carbonate as the best material combination of those evaluated. The program has successfully demonstrated techniques for folding large area systems into compact geometry's and vacuum impregnating the laminate in the folded systems. The major operational challenges encountered revolve around edge grading and low inductance, low impedance switching. The design iterations and lessons learned are discussed. A multistage prototype testing program has demonstrated 600kV operation on a short 6ns line. Full-scale prototypes are currently undergoing development and testing.

  1. Mutant lamin A links prophase to a p53 independent senescence program

    PubMed Central

    Moiseeva, Olga; Lessard, Frédéric; Acevedo-Aquino, Mariana; Vernier, Mathieu; Tsantrizos, Youla S; Ferbeyre, Gerardo

    2015-01-01

    Expression of oncogenes or short telomeres can trigger an anticancer response known as cellular senescence activating the p53 and RB tumor suppressor pathways. This mechanism is switched off in most tumor cells by mutations in p53 and RB signaling pathways. Surprisingly, p53 disabled tumor cells could be forced into senescence by expression of a mutant allele of the nuclear envelope protein lamin A. The pro-senescence lamin A mutant contains a deletion in the sequence required for processing by the protease ZMPSTE24 leading to accumulation of farnesylated lamin A in the nuclear envelope. In addition, the serine at position 22, a target for CDK1-dependent phosphorylation, was mutated to alanine, preventing CDK1-catalyzed nuclear envelope disassembly. The accumulation of this mutant lamin A compromised prophase to prometaphase transition leading to invaginations of the nuclear lamina, nuclear fragmentation and impaired chromosome condensation. Cells exited this impaired mitosis without cytokinesis and re-replicated their DNA ultimately arresting in interphase as polyploid cells with features of cellular senescence including increased expression of inflammatory gene products and a significant reduction of tumorigenicity in vivo. PMID:26029982

  2. Postbuckling of laminated anisotropic panels

    NASA Technical Reports Server (NTRS)

    Jeffrey, Glenda L.

    1987-01-01

    A two-part study of the buckling and postbuckling of laminated anisotropic plates with bending-extensional coupling is presented. The first part involves the development and application of a modified Rayleigh-Ritz analysis technique. Modifications made to the classical technique can be grouped into three areas. First, known symmetries of anisotropic panels are exploited in the selection of approximation functions. Second, a reduced basis technique based on these same symmetries is applied in the linear range. Finally, geometric boundary conditions are enforced via an exterior penalty function approach, rather than relying on choice of approximation functions to satisfy these boundary conditions. Numerical results are presented for both the linear and nonlinear range, with additional studies made to determine the effect of variation in penalty parameter and number of basis vectors. In the second part, six panels possessing anisotropy and bending-extensional coupling are tested. Detailed comparisons are made between experiment and finite element results in order to gain insight into the postbuckling and failure characteristics of such panels. The panels are constructed using two different lamination sequences, and panels with three different aspect ratios were constructed for each lamination sequence.

  3. Influence of Lamin A on the Mechanical Properties of Amphibian Oocyte Nuclei Measured by Atomic Force Microscopy

    PubMed Central

    Schäpe, Jens; Prauße, Steffi; Radmacher, Manfred; Stick, Reimer

    2009-01-01

    The nuclear lamina is part of the nuclear envelope (NE). Lamin filaments provide the nucleus with mechanical stability and are involved in many nuclear activities. The functional importance of these proteins is highlighted by mutations in lamin genes, which cause a variety of human diseases (laminopathies). Here we describe a method that allows one to quantify the contribution of lamin A protein to the mechanical properties of the NE. Lamin A is ectopically expressed in Xenopus oocytes, where it is incorporated into the NE of the oocyte nucleus, giving rise to a prominent lamina layer at the inner nuclear membrane. Nuclei are then isolated and probed by atomic force microscopy. From the resulting force curves, stiffness values are calculated and compared with those of control nuclei. Expression of lamin A significantly increases the stiffness of oocyte nuclei in a concentration-dependent manner. Since chromatin adds negligibly to nuclear mechanics in these giant nuclei, this method allows one to measure the contribution of individual NE components to nuclear mechanics. PMID:19450502

  4. 78 FR 48903 - Certain Products Having Laminated Packaging, Laminated Packaging, and Components Thereof...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-12

    ... From the Federal Register Online via the Government Publishing Office INTERNATIONAL TRADE COMMISSION Certain Products Having Laminated Packaging, Laminated Packaging, and Components Thereof... & Spirits Group of Cognac, France (``Camus''). Camus, Sidney Frank, and L'Oreal have since been...

  5. Novel origin of lamin-derived cytoplasmic intermediate filaments in tardigrades.

    PubMed

    Hering, Lars; Bouameur, Jamal-Eddine; Reichelt, Julian; Magin, Thomas M; Mayer, Georg

    2016-02-03

    Intermediate filament (IF) proteins, including nuclear lamins and cytoplasmic IF proteins, are essential cytoskeletal components of bilaterian cells. Despite their important role in protecting tissues against mechanical force, no cytoplasmic IF proteins have been convincingly identified in arthropods. Here we show that the ancestral cytoplasmic IF protein gene was lost in the entire panarthropod (onychophoran + tardigrade + arthropod) rather than arthropod lineage and that nuclear, lamin-derived proteins instead acquired new cytoplasmic roles at least three times independently in collembolans, copepods, and tardigrades. Transcriptomic and genomic data revealed three IF protein genes in the tardigrade Hypsibius dujardini, one of which (cytotardin) occurs exclusively in the cytoplasm of epidermal and foregut epithelia, where it forms belt-like filaments around each epithelial cell. These results suggest that a lamin derivative has been co-opted to enhance tissue stability in tardigrades, a function otherwise served by cytoplasmic IF proteins in all other bilaterians.

  6. Mechanisms of nuclear lamina growth in interphase.

    PubMed

    Zhironkina, Oxana A; Kurchashova, Svetlana Yu; Pozharskaia, Vasilisa A; Cherepanynets, Varvara D; Strelkova, Olga S; Hozak, Pavel; Kireev, Igor I

    2016-04-01

    The nuclear lamina represents a multifunctional platform involved in such diverse yet interconnected processes as spatial organization of the genome, maintenance of mechanical stability of the nucleus, regulation of transcription and replication. Most of lamina activities are exerted through tethering of lamina-associated chromatin domains (LADs) to the nuclear periphery. Yet, the lamina is a dynamic structure demonstrating considerable expansion during the cell cycle to accommodate increased number of LADs formed during DNA replication. We analyzed dynamics of nuclear growth during interphase and changes in lamina structure as a function of cell cycle progression. The nuclear lamina demonstrates steady growth from G1 till G2, while quantitative analysis of lamina meshwork by super-resolution microscopy revealed that microdomain organization of the lamina is maintained, with lamin A and lamin B microdomain periodicity and interdomain gap sizes unchanged. FRAP analysis, in contrast, demonstrated differences in lamin A and B1 exchange rates; the latter showing higher recovery rate in S-phase cells. In order to further analyze the mechanism of lamina growth in interphase, we generated a lamina-free nuclear envelope in living interphase cells by reversible hypotonic shock. The nuclear envelope in nuclear buds formed after such a treatment initially lacked lamins, and analysis of lamina formation revealed striking difference in lamin A and B1 assembly: lamin A reassembled within 30 min post-treatment, whereas lamin B1 did not incorporate into the newly formed lamina at all. We suggest that in somatic cells lamin B1 meshwork growth is coordinated with replication of LADs, and lamin A meshwork assembly seems to be chromatin-independent process.

  7. Glucocorticoids and laminitis in the horse.

    PubMed

    Johnson, Philip J; Slight, Simon H; Ganjam, Venkataseshu K; Kreeger, John M

    2002-08-01

    The administration of exogenously administered GCs and syndromes associated with GC excess are both attended by increased risk for the development of laminitis in adult horses. However, there exists substantial controversy as to whether excess GCs cause laminitis de novo. If true, the pathogenesis of laminitis arising from the effects of GC excess is probably different from that associated with diseases of the gastrointestinal tract and endotoxemia. Although a satisfactory explanation for the development of laminitis as a consequence of GC action is currently lacking, numerous possible and plausible theoretical mechanisms do exist. Veterinarians must exert caution with respect to the use of GCs in adult horses. The extent to which individual horses are predisposed to laminitis as a result of GC effect cannot be predicted based on current information. However, the administration of systemic GCs to horses that have been previously affected by laminitis should be used only with extreme caution, and should be accompanied by careful monitoring for further signs of laminitis. The risk of laminitis appears to be greater during treatment using some GCs (especially dexamethasone and triamcinalone) compared with others (prednisone and prednisolone). Whenever possible, to reduce the risk of laminitis, GCs should be administered locally. For example, the risk of GC-associated laminitis is evidently considerably reduced in horses affected with chronic obstructive pulmonary disease (COPD) if GC treatment is administered via inhalation. We have hypothesized that structural changes in the equine hoof that resemble laminitis may arise as a consequence of excess GC effect. Although these changes are not painful per se, and are not associated with inflammation, they could likely predispose affected horses to the development of bona fide laminitis for other reasons. Moreover, the gross morphological appearance of the chronically GC-affected hoof resembles that of a chronically

  8. The pathophysiology of developmental and acute laminitis.

    PubMed

    Hood, D M

    1999-08-01

    This review implies that although we know more regarding the enigma of developmental and acute laminitis today than previously, there is still more to investigate. As these investigations are conducted and interpreted, new and more effective preventive and therapeutic regimens are likely to be developed, tested, and made available. As this occurs, the impact of laminitis should undoubtedly decrease. Unfortunately, due to the lack of clinical symptoms in the developmental phase and the shortness of the acute phase, it is also evident that the two sequelae of acute laminitis, subacute and chronic laminitis, are likely to continue to pose a major problem for some time.

  9. Geometrically nonlinear analysis of laminated elastic structures

    NASA Technical Reports Server (NTRS)

    Reddy, J. N.

    1984-01-01

    Laminated composite plates and shells that can be used to model automobile bodies, aircraft wings and fuselages, and pressure vessels among many other were analyzed. The finite element method, a numerical technique for engineering analysis of structures, is used to model the geometry and approximate the solution. Various alternative formulations for analyzing laminated plates and shells are developed and their finite element models are tested for accuracy and economy in computation. These include the shear deformation laminate theory and degenerated 3-D elasticity theory for laminates.

  10. The laminA/NF-Y protein complex reveals an unknown transcriptional mechanism on cell proliferation.

    PubMed

    Cicchillitti, Lucia; Manni, Isabella; Mancone, Carmine; Regazzo, Giulia; Spagnuolo, Manuela; Alonzi, Tonino; Carlomosti, Fabrizio; Dell'Anna, Maria Lucia; Dell'Omo, Giulia; Picardo, Mauro; Ciana, Paolo; Capogrossi, Maurizio C; Tripodi, Marco; Magenta, Alessandra; Rizzo, Maria Giulia; Gurtner, Aymone; Piaggio, Giulia

    2017-01-10

    Lamin A is a component of the nuclear matrix that also controls proliferation by largely unknown mechanisms. NF-Y is a ubiquitous protein involved in cell proliferation composed of three subunits (-YA -YB -YC) all required for the DNA binding and transactivation activity. To get clues on new NF-Y partner(s) we performed a mass spectrometry screening of proteins that co-precipitate with the regulatory subunit of the complex, NF-YA. By this screening we identified lamin A as a novel putative NF-Y interactor. Co-immunoprecipitation experiments and confocal analysis confirmed the interaction between the two endogenous proteins. Interestingly, this association occurs on euchromatin regions, too. ChIP experiments demonstrate lamin A enrichment in several promoter regions of cell cycle related genes in a NF-Y dependent manner. Gain and loss of function experiments reveal that lamin A counteracts NF-Y transcriptional activity. Taking advantage of a recently generated transgenic reporter mouse, called MITO-Luc, in which an NF-Y-dependent promoter controls luciferase expression, we demonstrate that lamin A counteracts NF-Y transcriptional activity not only in culture cells but also in living animals. Altogether, our data demonstrate the occurrence of lamin A/NF-Y interaction and suggest a possible role of this protein complex in regulation of NF-Y function in cell proliferation.

  11. Lamin A/C Haploinsufficiency Causes Dilated Cardiomyopathy and Apoptosis-Triggered Cardiac Conduction System Disease

    PubMed Central

    Wolf, Cordula M; Wang, Libin; Alcalai, Ronny; Pizard, Anne; Burgon, Patrick G; Ahmad, Ferhaan; Sherwood, Megan; Branco, Dorothy M; Wakimoto, Hiroko; Fishman, Glenn I; See, Vincent; Stewart, Colin L; Conner, David A; Berul, Charles I; Seidman, Christine E; Seidman, JG

    2008-01-01

    Mutations in the lamin A/C (LMNA) gene, which encodes nuclear membrane proteins, cause a variety of human conditions including dilated cardiomyopathy (DCM) with associated cardiac conduction system disease. To investigate mechanisms responsible for electrophysiologic and myocardial phenotypes caused by dominant human LMNA mutations, we performed longitudinal evaluations in heterozygous Lmna+/- mice. Despite one normal allele, Lmna+/- mice had 50% of normal cardiac lamin A/C levels and developed cardiac abnormalities. Conduction system function was normal in neonatal Lmna+/- mice but by 4 weeks of age AV nodal myocytes had abnormally shaped nuclei and active apoptosis. Telemetric and in vivo electrophysiologic studies in 10 week-old Lmna+/- mice showed atrioventricular (AV) conduction defects and both atrial and ventricular arrhythmias, analogous to those observed in humans with heterozygous LMNA mutations. Isolated myocytes from 12-month old Lmna+/- mice exhibited impaired contractility. In vivo cardiac studies of aged Lmna+/- mice revealed DCM; in some mice this occurred without overt conduction system disease. However, neither histopathology nor serum CK levels indicated skeletal muscle pathology. These data demonstrate cardiac pathology due to heterozygous Lmna mutations reflecting a 50% reduction in lamin protein levels. Lamin haploinsufficiency caused early-onset programmed cell death of AV nodal myocytes and progressive electrophysiologic disease. While lamin haploinsufficiency was better tolerated by non-conducting myocytes, ultimately these too succumbed to diminished lamin levels leading to dilated cardiomyopathy, which presumably arose independently from conduction system disease. PMID:18182166

  12. Lamin-like analogues in plants: the characterization of NMCP1 in Allium cepa

    PubMed Central

    Moreno Díaz de la Espina, Susana

    2013-01-01

    The nucleoskeleton of plants contains a peripheral lamina (also called plamina) and, even though lamins are absent in plants, their roles are still fulfilled in plant nuclei. One of the most intriguing topics in plant biology concerns the identity of lamin protein analogues in plants. Good candidates to play lamin functions in plants are the members of the NMCP (nuclear matrix constituent protein) family, which exhibit the typical tripartite structure of lamins. This paper describes a bioinformatics analysis and classification of the NMCP family based on phylogenetic relationships, sequence similarity and the distribution of conserved regions in 76 homologues. In addition, NMCP1 in the monocot Allium cepa characterized by its sequence and structure, biochemical properties, and subnuclear distribution and alterations in its expression throughout the root were identified. The results demonstrate that these proteins exhibit many similarities to lamins (structural organization, conserved regions, subnuclear distribution, and solubility) and that they may fulfil the functions of lamins in plants. These findings significantly advance understanding of the structural proteins of the plant lamina and nucleoskeleton and provide a basis for further investigation of the protein networks forming these structures. PMID:23378381

  13. Field performance of stress-laminated highway bridges constructed with glued laminated timber

    Treesearch

    J.P. Wacker

    2004-01-01

    This paper summarizes the field performance of three stress-laminated deck timber bridges located in Wisconsin, New York, and Arizona. The deck superstructures of these single-span highway bridges is comprised of full-span glued laminated timber (glulam) beam laminations manufactured with southern pine, hem fir/red maple combination, and/or Douglas fir lumber species....

  14. Nuclear membrane protein emerin: roles in gene regulation, actin dynamics and human disease.

    PubMed

    Wilson, Katherine L; Holaska, James M; Montes de Oca, Rocio; Tifft, Kathryn; Zastrow, Michael; Segura-Totten, Miriam; Mansharamani, Malini; Bengtsson, Luiza

    2005-01-01

    Loss of emerin, a nuclear membrane protein, causes Emery-Dreifuss muscular dystrophy (EDMD), characterized by muscle weakening, contractures of major tendons and potentially lethal cardiac conduction system defects. Emerin has a LEM-domain and therefore binds barrier-to-autointegration factor (BAF), a conserved chromatin protein essential for cell division. BAF recruits emerin to chromatin and regulates higher-order chromatin structure during nuclear assembly. Emerin also binds filaments formed by A-type lamins, mutations in which also cause EDMD. Other partners for emerin include nesprin-1alpha and transcriptional regulators such as germ cell-less (GCL). The binding affinities of these partners range from 4nM (nesprin-1alpha) to 200 nM (BAF), and are physiologically significant. Biochemical studies therefore provide a valid means to predict the properties of emerin-lamin complexes in vivo. Emerin and lamin A together form stable complexes with either BAF or GCL in vitro. BAF, however, competes with GCL for binding to emerin in vitro. These and additional partners, notably actin and nuclear myosin II, suggest disease-relevant roles for emerin in gene regulation and the mechanical interity of the nucleus.

  15. Methods to Monitor DNA Repair Defects and Genomic Instability in the Context of a Disrupted Nuclear Lamina

    PubMed Central

    Gonzalo, Susana; Kreienkamp, Ray

    2016-01-01

    The organization of the genome within the nuclear space is viewed as an additional level of regulation of genome function, as well as a means to ensure genome integrity. Structural proteins associated with the nuclear envelope, in particular lamins (A- and B-type) and lamin-associated proteins, play an important role in genome organization. Interestingly, there is a whole body of evidence that links disruptions of the nuclear lamina with DNA repair defects and genomic instability. Here, we describe a few standard techniques that have been successfully utilized to identify mechanisms behind DNA repair defects and genomic instability in cells with an altered nuclear lamina. In particular, we describe protocols to monitor changes in the expression of DNA repair factors (Western blot) and their recruitment to sites of DNA damage (immunofluorescence); kinetics of DNA double-strand break repair after ionizing radiation (neutral comet assays); frequency of chromosomal aberrations (FISH, fluorescence in situ hybridization); and alterations in telomere homeostasis (Quantitative-FISH). These techniques have allowed us to shed some light onto molecular mechanisms by which alterations in A-type lamins induce genomic instability, which could contribute to the pathophysiology of aging and aging-related diseases. PMID:27147057

  16. Autophagy mediates degradation of nuclear lamina

    PubMed Central

    Dou, Zhixun; Xu, Caiyue; Donahue, Greg; Shimi, Takeshi; Pan, Ji-An; Zhu, Jiajun; Ivanov, Andrejs; Capell, Brian C.; Drake, Adam M.; Shah, Parisha P.; Catanzaro, Joseph M.; Ricketts, M. Daniel; Lamark, Trond; Adam, Stephen A.; Marmorstein, Ronen; Zong, Wei-Xing; Johansen, Terje; Goldman, Robert D.; Adams, Peter D.; Berger, Shelley L.

    2015-01-01

    Macroautophagy (hereafter referred to as autophagy) is a catabolic membrane trafficking process that degrades a variety of cellular constituents, and is associated with human diseases1–3. While extensive studies have focused on autophagic turnover of cytoplasmic materials, little is known regarding the role of autophagy in degrading nuclear components. Here we report that the autophagy machinery mediates degradation of nuclear lamina components in mammals. The autophagy protein LC3/Atg8, which is involved in autophagy membrane trafficking and substrate delivery4–6, is present in the nucleus and directly interacts with the nuclear lamina protein Lamin B1, and binds to lamin-associated domains (LADs) on chromatin. This LC3-Lamin B1 interaction does not downregulate Lamin B1 during starvation, but mediates its degradation upon oncogenic insults, such as by activated Ras. Lamin B1 degradation is achieved by nucleus-to-cytoplasm transport that delivers Lamin B1 to the lysosome. Inhibiting autophagy or the LC3-Lamin B1 interaction prevents activated Ras-induced Lamin B1 loss and attenuates oncogene-induced senescence in primary human cells. Our study suggests this new function of autophagy as a guarding mechanism protecting cells from tumorigenesis. PMID:26524528

  17. Autophagy mediates degradation of nuclear lamina.

    PubMed

    Dou, Zhixun; Xu, Caiyue; Donahue, Greg; Shimi, Takeshi; Pan, Ji-An; Zhu, Jiajun; Ivanov, Andrejs; Capell, Brian C; Drake, Adam M; Shah, Parisha P; Catanzaro, Joseph M; Ricketts, M Daniel; Lamark, Trond; Adam, Stephen A; Marmorstein, Ronen; Zong, Wei-Xing; Johansen, Terje; Goldman, Robert D; Adams, Peter D; Berger, Shelley L

    2015-11-05

    Macroautophagy (hereafter referred to as autophagy) is a catabolic membrane trafficking process that degrades a variety of cellular constituents and is associated with human diseases. Although extensive studies have focused on autophagic turnover of cytoplasmic materials, little is known about the role of autophagy in degrading nuclear components. Here we report that the autophagy machinery mediates degradation of nuclear lamina components in mammals. The autophagy protein LC3/Atg8, which is involved in autophagy membrane trafficking and substrate delivery, is present in the nucleus and directly interacts with the nuclear lamina protein lamin B1, and binds to lamin-associated domains on chromatin. This LC3-lamin B1 interaction does not downregulate lamin B1 during starvation, but mediates its degradation upon oncogenic insults, such as by activated RAS. Lamin B1 degradation is achieved by nucleus-to-cytoplasm transport that delivers lamin B1 to the lysosome. Inhibiting autophagy or the LC3-lamin B1 interaction prevents activated RAS-induced lamin B1 loss and attenuates oncogene-induced senescence in primary human cells. Our study suggests that this new function of autophagy acts as a guarding mechanism protecting cells from tumorigenesis.

  18. Characterization of NE81, the first lamin-like nucleoskeleton protein in a unicellular organism.

    PubMed

    Krüger, Anne; Batsios, Petros; Baumann, Otto; Luckert, Eva; Schwarz, Heinz; Stick, Reimer; Meyer, Irene; Gräf, Ralph

    2012-01-01

    Lamins build the nuclear lamina and are required for chromatin organization, gene expression, cell cycle progression, and mechanical stabilization. Despite these universal functions, lamins have so far been found only in metazoans. We have identified protein NE81 in Dictyostelium, which has properties that justify its denomination as a lamin-like protein in a lower eukaryote. This is based on its primary structure, subcellular localization, and regulation during mitosis, and its requirement of the C-terminal CaaX box as a posttranslational processing signal for proper localization. Our knockout and overexpression mutants revealed an important role for NE81 in nuclear integrity, chromatin organization, and mechanical stability of cells. All our results are in agreement with a role for NE81 in formation of a nuclear lamina. This function is corroborated by localization of Dictyostelium NE81 at the nuclear envelope in human cells. The discovery of a lamin-like protein in a unicellular organism is not only intriguing in light of evolution, it may also provide a simple experimental platform for studies of the molecular basis of laminopathies.

  19. Lamin A/C sustains PcG protein architecture, maintaining transcriptional repression at target genes

    PubMed Central

    Cesarini, Elisa; Mozzetta, Chiara; Marullo, Fabrizia; Gregoretti, Francesco; Gargiulo, Annagiusi; Columbaro, Marta; Cortesi, Alice; Antonelli, Laura; Di Pelino, Simona; Squarzoni, Stefano; Palacios, Daniela; Zippo, Alessio; Bodega, Beatrice; Oliva, Gennaro

    2015-01-01

    Beyond its role in providing structure to the nuclear envelope, lamin A/C is involved in transcriptional regulation. However, its cross talk with epigenetic factors—and how this cross talk influences physiological processes—is still unexplored. Key epigenetic regulators of development and differentiation are the Polycomb group (PcG) of proteins, organized in the nucleus as microscopically visible foci. Here, we show that lamin A/C is evolutionarily required for correct PcG protein nuclear compartmentalization. Confocal microscopy supported by new algorithms for image analysis reveals that lamin A/C knock-down leads to PcG protein foci disassembly and PcG protein dispersion. This causes detachment from chromatin and defects in PcG protein–mediated higher-order structures, thereby leading to impaired PcG protein repressive functions. Using myogenic differentiation as a model, we found that reduced levels of lamin A/C at the onset of differentiation led to an anticipation of the myogenic program because of an alteration of PcG protein–mediated transcriptional repression. Collectively, our results indicate that lamin A/C can modulate transcription through the regulation of PcG protein epigenetic factors. PMID:26553927

  20. Interruption of progerin–lamin A/C binding ameliorates Hutchinson-Gilford progeria syndrome phenotype

    PubMed Central

    Lee, Su-Jin; Jung, Youn-Sang; Yoon, Min-Ho; Kang, So-mi; Oh, Ah-Young; Lee, Jee-Hyun; Jun, So-Young; Woo, Tae-Gyun; Chun, Ho-Young; Kim, Sang Kyum; Chung, Kyu Jin; Lee, Ho-Young; Lee, Kyeong; Jin, Guanghai; Na, Min-Kyun; Ha, Nam Chul; Bárcena, Clea; Freije, José M.P.; López-Otín, Carlos; Song, Gyu Yong

    2016-01-01

    Hutchinson-Gilford progeria syndrome (HGPS) is a rare autosomal dominant genetic disease that is caused by a silent mutation of the LMNA gene encoding lamins A and C (lamin A/C). The G608G mutation generates a more accessible splicing donor site than does WT and produces an alternatively spliced product of LMNA called progerin, which is also expressed in normal aged cells. In this study, we determined that progerin binds directly to lamin A/C and induces profound nuclear aberrations. Given this observation, we performed a random screening of a chemical library and identified 3 compounds (JH1, JH4, and JH13) that efficiently block progerin–lamin A/C binding. These 3 chemicals, particularly JH4, alleviated nuclear deformation and reversed senescence markers characteristic of HGPS cells, including growth arrest and senescence-associated β-gal (SA–β-gal) activity. We then used microarray-based analysis to demonstrate that JH4 is able to rescue defects of cell-cycle progression in both HGPS and aged cells. Furthermore, administration of JH4 to LmnaG609G/G609G-mutant mice, which phenocopy human HGPS, resulted in a marked improvement of several progeria phenotypes and an extended lifespan. Together, these findings indicate that specific inhibitors with the ability to block pathological progerin–lamin A/C binding may represent a promising strategy for improving lifespan and health in both HGPS and normal aging. PMID:27617860

  1. Interruption of progerin-lamin A/C binding ameliorates Hutchinson-Gilford progeria syndrome phenotype.

    PubMed

    Lee, Su-Jin; Jung, Youn-Sang; Yoon, Min-Ho; Kang, So-Mi; Oh, Ah-Young; Lee, Jee-Hyun; Jun, So-Young; Woo, Tae-Gyun; Chun, Ho-Young; Kim, Sang Kyum; Chung, Kyu Jin; Lee, Ho-Young; Lee, Kyeong; Jin, Guanghai; Na, Min-Kyun; Ha, Nam Chul; Bárcena, Clea; Freije, José M P; López-Otín, Carlos; Song, Gyu Yong; Park, Bum-Joon

    2016-10-03

    Hutchinson-Gilford progeria syndrome (HGPS) is a rare autosomal dominant genetic disease that is caused by a silent mutation of the LMNA gene encoding lamins A and C (lamin A/C). The G608G mutation generates a more accessible splicing donor site than does WT and produces an alternatively spliced product of LMNA called progerin, which is also expressed in normal aged cells. In this study, we determined that progerin binds directly to lamin A/C and induces profound nuclear aberrations. Given this observation, we performed a random screening of a chemical library and identified 3 compounds (JH1, JH4, and JH13) that efficiently block progerin-lamin A/C binding. These 3 chemicals, particularly JH4, alleviated nuclear deformation and reversed senescence markers characteristic of HGPS cells, including growth arrest and senescence-associated β-gal (SA-β-gal) activity. We then used microarray-based analysis to demonstrate that JH4 is able to rescue defects of cell-cycle progression in both HGPS and aged cells. Furthermore, administration of JH4 to LmnaG609G/G609G-mutant mice, which phenocopy human HGPS, resulted in a marked improvement of several progeria phenotypes and an extended lifespan. Together, these findings indicate that specific inhibitors with the ability to block pathological progerin-lamin A/C binding may represent a promising strategy for improving lifespan and health in both HGPS and normal aging.

  2. Lamin A/C sustains PcG protein architecture, maintaining transcriptional repression at target genes.

    PubMed

    Cesarini, Elisa; Mozzetta, Chiara; Marullo, Fabrizia; Gregoretti, Francesco; Gargiulo, Annagiusi; Columbaro, Marta; Cortesi, Alice; Antonelli, Laura; Di Pelino, Simona; Squarzoni, Stefano; Palacios, Daniela; Zippo, Alessio; Bodega, Beatrice; Oliva, Gennaro; Lanzuolo, Chiara

    2015-11-09

    Beyond its role in providing structure to the nuclear envelope, lamin A/C is involved in transcriptional regulation. However, its cross talk with epigenetic factors--and how this cross talk influences physiological processes--is still unexplored. Key epigenetic regulators of development and differentiation are the Polycomb group (PcG) of proteins, organized in the nucleus as microscopically visible foci. Here, we show that lamin A/C is evolutionarily required for correct PcG protein nuclear compartmentalization. Confocal microscopy supported by new algorithms for image analysis reveals that lamin A/C knock-down leads to PcG protein foci disassembly and PcG protein dispersion. This causes detachment from chromatin and defects in PcG protein-mediated higher-order structures, thereby leading to impaired PcG protein repressive functions. Using myogenic differentiation as a model, we found that reduced levels of lamin A/C at the onset of differentiation led to an anticipation of the myogenic program because of an alteration of PcG protein-mediated transcriptional repression. Collectively, our results indicate that lamin A/C can modulate transcription through the regulation of PcG protein epigenetic factors.

  3. Atomic force microscopy and lamins: A review study towards future, combined investigations.

    PubMed

    Pecorari, Ilaria; Puzzi, Luca; Sbaizero, Orfeo

    2017-01-01

    In the last decades, atomic force microscopy (AFM) underwent a rapid and stunning development, especially for studying mechanical properties of biological samples. The numerous discoveries relying to this approach, have increased the credit of AFM as a versatile tool, and potentially eligible as a diagnostic equipment. Meanwhile, it has become strikingly evident that lamins are involved on the onset and development of certain diseases, including cancer, Hutchinson-Gilford progeria syndrome, cardiovascular pathologies, and muscular dystrophy. A new category of pathologies has been defined, the laminopathies, which are caused by mutations in the gene encoding for A-type lamins. As the majority of medical issues, lamins, and all their related aspects can be considered as a quite complex problem. Indeed, there are many facets to explore, and this definitely requires a multidisciplinary approach. One of the most intriguing aspects concerning lamins is their remarkable contribute to cells mechanics. Over the years, this has led to the speculation of the so-called "structural hypothesis", which attempts to elucidate the etiology and some features of the laminopathies. Among the various techniques tried to figure out the role of lamins in the cells mechanics, the AFM has been already successfully applied, proving its versatility. Therefore, the present work aims both to highlight the qualities of AFM and to review the most relevant knowledge about lamins, in order to promote the study of the latter, taking advantage from the former. Microsc. Res. Tech. 80:97-108, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  4. Machining of fiber-reinforced composite laminates

    NASA Astrophysics Data System (ADS)

    Won, Myong-Shik

    As fiber-reinforced composite laminates are becoming considerably popular in a wide range of applications, the necessity for machining such materials is increasing rapidly. Due to their microscopical inhomogeneity, anisotropy, and highly abrasive nature, composite laminates exhibit some peculiar types of machining damage. Consequently, the machining of composite laminates requires a different approach from that used for metals and offers a challenge from both an academic and application point of view. In the present work, the drilling of composite laminated plates and the edge trimming of tubular composite laminates were investigated through theoretical analyses and their experimental verification. First, a drilling process model using linear elastic fracture mechanics and classical plate bending theory was developed to predict the critical thrust value responsible for the onset of delamination during the drilling of composite laminates with pre-drilled pilot holes. Experiments using stepped drills, which can utilize the effectiveness of such pilot holes, were conducted on composite laminates. Reasonably good agreement was found between the results of the process model and the tests. Second, the development of a model-based intelligent control strategy for the efficient drilling of composite laminates was explored by experiments and analyses. In this investigation, mathematical models were created to relate the drilling forces to cutting parameters and to identify the different process stages. These models predicted the degree of thrust force regulation to prevent delamination. Third, the edge trimming of thin-walled tubular composite laminates was modeled and analyzed for estimating the critical cutting force at the initiation of longitudinal cracking. A series of full-scale edge trimming tests were conducted on tubular composite specimens to assess the current approach and to obtain basic machining data for various composite laminates. The present study provides

  5. Effect of laminate thickness and specimen configuration on the fracture of laminated composites

    NASA Technical Reports Server (NTRS)

    Harris, C. E.; Morris, D. H.

    1986-01-01

    Attention is given to the effect of laminate thickness on graphite/epoxy laminates in the present measurements of fracture toughness in center cracked tension specimens, compact tension specimens, and three-point bend specimens. Crack tip damage development prior to fracture is also studied. The results obtained show fracture toughness to be a function of laminate thickness, being in all cases independent of crack size. The fracture surface of all thick laminates was uniform in the interior and self-similar with the starter notch. With only one exception, the fracture toughness of the thicker laminates was relatively independent of specimen configuration.

  6. Negative refraction in a laminate

    NASA Astrophysics Data System (ADS)

    Willis, J. R.

    2016-12-01

    This work is concerned with the reflection and transmission of waves at a plane interface between a homogeneous elastic half-space and a half-space of elastic material that is periodically laminated. The lamination is always in the direction of the x1-coordinate axis and the displacement is always longitudinal shear, so that the only non-zero displacement component is u3(x1 ,x2 , t). After an initial discussion of Floquet-Bloch waves in the laminated material, brief consideration is given to the reflection-transmission problem, when the interface between the two media is the plane x1 = 0. Nothing unusual emerges: there are just a single reflected wave and a single transmitted wave, undergoing positive group-velocity refraction. Then, the problem is considered when the interface between the two media is the plane x2 = 0. The periodic structure of the interface induces an infinite set of reflected waves and an infinite set of transmitted waves. All need to be taken into account, but most decay exponentially away from the interface. It had previously been recognized that, if the incident wave had appropriate frequency and angle of incidence, a propagating transmitted wave would be generated that would undergo negative group-velocity refraction - behaviour usually associated with a metamaterial. It is established by an example in this work that there is, in addition, a propagating transmitted wave with smaller wavelength but larger group velocity that undergoes positive group-velocity refraction. The work concludes with a brief discussion of this finding, including its implications for the utility (or not) of "effective medium" theory.

  7. Symmetries in laminated composite plates

    NASA Technical Reports Server (NTRS)

    Noor, A. K.

    1976-01-01

    The different types of symmetry exhibited by laminated anisotropic fibrous composite plates are identified and contrasted with the symmetries of isotropic and homogeneous orthotropic plates. The effects of variations in the fiber orientation and the stacking sequence of the layers on the symmetries exhibited by composite plates are discussed. Both the linear and geometrically nonlinear responses of the plates are considered. A simple procedure is presented for exploiting the symmetries in the finite element analysis. Examples are given of square, skew and polygonal plates where use of symmetry concepts can significantly reduce the scope and cost of analysis.

  8. Maggot debridement therapy for laminitis.

    PubMed

    Morrison, Scott

    2010-08-01

    Maggot debridement therapy is a nontraumatic, minimally invasive method to treat infections in a foot compromised by chronic laminitis. A mechanical strategy must first be in place to address the instability of the distal phalanx and hoof capsule. Adverse reactions to maggot debridement therapy are uncommon and the only side effect observed has been irritation or hypersensitivity at the site. Chronic laminitic cases of sepsis/necrosis within the hoof benefit from this procedure due to the noninvasive, continuous debridement and healing properties provided by the larvae.

  9. Residual stresses in angleplied laminates and their effects on laminate behavior

    NASA Technical Reports Server (NTRS)

    Chamis, C. C.

    1978-01-01

    Evidence of the presence of lamination residual stresses in angleplied laminates were transply cracks and warpage of unsymmetric laminates which occur prior to application of any mechanical load. Lamination residual strains were measured using the embedded strain gage technique. These strains result from the temperature differences between cure and room temperature and vary linearly within this temperature range. Lamination residual stresses were usually present in angleplied fiber composites laminates; they were also present in unidirectional hybrids and superhybrids. For specific applications, the magnitudes of lamination residual stresses were determined and evaluated relative to the anticipated applied stresses. Particular attention was given to cyclic thermal loadings in applications where the thermal cycling takes place over a wide temperature range.

  10. Evaluating the warping of laminated particleboard panels

    Treesearch

    Zhiyong Cai

    2004-01-01

    Laminated wood composites have been used widely in the secondary manufacturing processes in the wood panel industries. Warping, which is defined as the out-of-plane deformation of an initially flat panel, is a longstanding problem associated with the use of laminated wood composites. The mechanism of warping is still not fully understood. A new two- dimensional warping...

  11. Silicone modified resins for graphite fiber laminates

    NASA Technical Reports Server (NTRS)

    Frost, L. W.; Bower, G. M.

    1980-01-01

    Six silicone modified resins were selected for evaluation in unidirectional filament wound graphite laminates. Neat samples of these resins had 1,000 C char residues of 6-63%. The highest flexural values measured for the laminates were a strength of 1,220 MPa and a modulus of 105 GPa. The highest interlaminar shear strength was 72 MPa.

  12. The strength of Norwegian glued laminated beams

    Treesearch

    Kjell Solli; Erik Aasheim; Robert H. Falk

    1992-01-01

    This paper focuses on the characterization and the performance of glued laminated (glulam) timber beams manufactured from machine stress graded Norwegian spruce in comparison to developing CEN standards. Material property testing indicated that the supplied laminating timber can be represented by two CEN strength classes, C37-14E and C30-12E, with about 50% yield in...

  13. Characterization of unfolding mechanism of human lamin A Ig fold by single-molecule force spectroscopy-implications in EDMD.

    PubMed

    Bera, Manindra; Kotamarthi, Hema Chandra; Dutta, Subarna; Ray, Angana; Ghosh, Saptaparni; Bhattacharyya, Dhananjay; Ainavarapu, Sri Rama Koti; Sengupta, Kaushik

    2014-11-25

    A- and B-type lamins are intermediate filament proteins constituting the nuclear lamina underneath the nuclear envelope thereby conferring proper shape and mechanical rigidity to the nucleus. Lamin proteins are also shown to be related diversely to basic nuclear processes. More than 400 mutations in human lamin A protein alone have been reported to produce at least 11 different disease conditions jointly termed as laminopathies. These mutations in lamin A are scattered throughout its helical rod domain, as well as the C-terminal domain containing the conserved Ig-fold region. The commonality of phenotypes in all these diseases is characterized by misshapen nuclei of the affected tissues which might stem from altered rigidity of the supporting lamina hence lamins. Here we have focused on autosomal dominant Emery-Dreifuss Muscular Dystrophy, one such laminopathy where R453W is the causative mutation located in the Ig domain of lamin A. We have investigated by single-molecule force spectroscopy how a stretching mechanical perturbation senses the destabilizing effect of the mutation in the lamin A Ig domain and compared the mechanoelastic properties of the mutant R453W with that of the wild-type in conjunction with steered molecular dynamics. Furthermore, we have shown the interaction of Ig domain with emerin, another key player and interacting partner in the pathogenesis of EDMD, is disrupted in the R453W mutant. This altered mechanoresistance of Ig domain itself and consequent uncoupling of lamin A-emerin interaction might underlie the altered mechanotransduction properties of EDMD affected nuclei.

  14. Stationary turbine component with laminated skin

    DOEpatents

    James, Allister W [Orlando, FL

    2012-08-14

    A stationary turbine engine component, such as a turbine vane, includes a internal spar and an external skin. The internal spar is made of a plurality of spar laminates, and the external skin is made of a plurality of skin laminates. The plurality of skin laminates interlockingly engage the plurality of spar laminates such that the external skin is located and held in place. This arrangement allows alternative high temperature materials to be used on turbine engine components in areas where their properties are needed without having to make the entire component out of such material. Thus, the manufacturing difficulties associated with making an entire component of such a material and the attendant high costs are avoided. The skin laminates can be made of advanced generation single crystal superalloys, intermetallics and refractory alloys.

  15. Wettability of graphene-laminated micropillar structures

    NASA Astrophysics Data System (ADS)

    Bong, Jihye; Seo, Keumyoung; Park, Ji-Hoon; Ahn, Joung Real; Ju, Sanghyun

    2014-12-01

    The wetting control of graphene is of great interest for electronic, mechanical, architectural, and bionic applications. In this study, the wettability of graphene-laminated micropillar structures was manipulated by changing the height of graphene-laminated structures and employing the trichlorosilane (HDF-S)-based self-assembly monolayer. Graphene-laminated micropillar structures with HDF-S exhibited higher hydrophobicity (contact angle of 129.5°) than pristine graphene thin film (78.8°), pristine graphene-laminated micropillar structures (97.5°), and HDF-S self-assembled graphene thin film (98.5°). Wetting states of the graphene-laminated micropillar structure with HDF-S was also examined by using a urea solution, which flowed across the surface without leaving any residues.

  16. Steady compression characteristics of laminated MRE isolator

    NASA Astrophysics Data System (ADS)

    Wahab, N. A. A.; Mazlan, S. A.; Ubaidillah; Sharif, A. H. R.; Kamaruddin, S.

    2016-11-01

    This paper focused on an experimental setup on laminated magnetorheological elastomer (MRE) isolator under steady state compression test. An isotropic type natural rubber (NR) based MRE were fabricated and layered with a steel plate to form a multilayer sandwich structure adopted from the conventional laminated rubber bearing design. A set of static compression test was conducted to explore the potential of semi-active laminated MRE isolator in field-dependent stiffness properties. Stress versus strain relationship was assessed under different magnetic fields application. Based on the examination, the stress altered as the application of magnetic fields. Consequently, the effective stiffness of isolator also influenced by the magnetic fields induction. The experimental results show that the proposed laminated MRE isolator can effectively alter the compression stiffness up to the 14.56%. The preliminary results have confirmed the tunability of the semi-active laminated MRE isolator in which it would be beneficial for improving building isolator in general.

  17. Universality of the frequency spectrum of laminates

    NASA Astrophysics Data System (ADS)

    Shmuel, Gal; Band, Ram

    2016-07-01

    We show that the frequency spectrum of two-component elastic laminates admits a universal structure, independent of the geometry of the periodic-cell and the specific physical properties. The compactness of the structure enables us to rigorously derive the maximal width, the expected width, and the density of the band-gaps - ranges of frequencies at which waves cannot propagate. In particular, we find that the density of these band-gaps is a universal property of classes of laminates. Rules for tailoring laminates according to desired spectrum properties thereby follow. We show that the frequency spectrum of various finitely deformed laminates are also endowed with the same compact structure. Finally, we explain how our results generalize for laminates with an arbitrary number of components, based on the form of their dispersion relation.

  18. Wettability of graphene-laminated micropillar structures

    SciTech Connect

    Bong, Jihye; Seo, Keumyoung; Ju, Sanghyun E-mail: shju@kgu.ac.kr; Park, Ji-Hoon; Ahn, Joung Real E-mail: shju@kgu.ac.kr

    2014-12-21

    The wetting control of graphene is of great interest for electronic, mechanical, architectural, and bionic applications. In this study, the wettability of graphene-laminated micropillar structures was manipulated by changing the height of graphene-laminated structures and employing the trichlorosilane (HDF-S)-based self-assembly monolayer. Graphene-laminated micropillar structures with HDF-S exhibited higher hydrophobicity (contact angle of 129.5°) than pristine graphene thin film (78.8°), pristine graphene-laminated micropillar structures (97.5°), and HDF-S self-assembled graphene thin film (98.5°). Wetting states of the graphene-laminated micropillar structure with HDF-S was also examined by using a urea solution, which flowed across the surface without leaving any residues.

  19. In Situ Detection of Interactions Between Nuclear Envelope Proteins and Partners.

    PubMed

    Barateau, Alice; Buendia, Brigitte

    2016-01-01

    Proximity ligation assay (PLA) appears as a quick and easy technique to visualize within fixed cells the occurrence and in situ distribution of protein complexes. PLA has been validated to detect protein-protein interactions within the nuclear compartment. Here, we describe a protocol which allows the detection of interactions between A-type nuclear lamins and either LEM-domain proteins (such as emerin, integrated within the inner nuclear membrane, and LAP2α which accumulates within the nucleoplasm) or gene regulatory factors (e.g., the transcription factor SREBP1). The distinct amounts and patterns of PLA signals obtained for various complexes highlight the pertinence of using PLA to reveal in situ where and to which extent nuclear envelope proteins bind specific partners.

  20. X-linked Emery-Dreifuss muscular dystrophy with lamin A deficiency and IBM inclusions.

    PubMed

    Fidzianska, A; Niebrój-Dobosz, I; Madej-Pilarczyk, A; Duong, N T; Wehnert, M

    2010-01-01

    The study demonstrates a 12-year-old patient with progressive proximal muscle weakness, joint contractures, rigidity of the neck, and absence of emerin and lamin A in the muscle nuclei, which is caused by intronic mutation IVS3-27del18 (c.266-27del18) in the emerin gene. The most surprising finding was the appearance of IBM-like inclusions in euchromatin, as well as aberrant nuclei. It may be speculated that altered expression of the emerin-lamin complex and modification of the nuclear matrix leads to formation of tubulofilamentous structures in the presented case.

  1. Mutation Glu82Lys in lamin A/C gene is associated with cardiomyopathy and conduction defect

    SciTech Connect

    Wang Hu; Wang Jizheng; Zheng Weiyue; Wang Xiaojian; Wang Shuxia; Song Lei; Zou Yubao; Yao Yan; Hui Rutai . E-mail: huirutai@sglab.org

    2006-05-26

    Dilated cardiomyopathy is a form of heart muscle disease characterized by impaired systolic function and ventricular dilation. The mutations in lamin A/C gene have been linked to dilated cardiomyopathy. We screened genetic mutations in a large Chinese family of 50 members including members with dilated cardiomyopathy and found a Glu82Lys substitution mutation in the rod domain of the lamin A/C protein in eight family members, three of them have been diagnosed as dilated cardiomyopathy, one presented with heart dilation. The pathogenic mechanism of lamin A/C gene defect is poorly understood. Glu82Lys mutated lamin A/C and wild type protein was transfected into HEK293 cells. The mutated protein was not properly localized at the inner nuclear membrane and the emerin protein, which interacts with lamin A/C, was also aberrantly distributed. The nuclear membrane structure was disrupted and heterochromatin was aggregated aberrantly in the nucleus of the HEK293 cells stably transfected with mutated lamin A/C gene as determined by transmission electron microscopy.

  2. Laminitis and the equine metabolic syndrome.

    PubMed

    Johnson, Philip J; Wiedmeyer, Charles E; LaCarrubba, Alison; Ganjam, V K Seshu; Messer, Nat T

    2010-08-01

    Although much has been written about laminitis in the context of its association with inflammatory processes, recognition is growing that most cases of laminitis examined by veterinarians in private practice are those associated with pasture grazing, obesity, and insulin resistance (IR). The term 'endocrinopathic laminitis' has been adopted to classify the instances of laminitis in which the origin seems to be more strongly associated with an underlying endocrinopathy, such as either IR or the influence of corticosteroids. Results of a recent study suggest that obesity and IR represent the most common metabolic and endocrinopathic predispositions for laminitis in horses. IR also plays an important role in the pathogenesis of laminitis that develops when some horses or ponies are allowed to graze pastures at certain times of the year. The term equine metabolic syndrome (EMS) has been proposed as a label for horses whose clinical examination results (including both physical examination and laboratory testing) suggest heightened risk for developing laminitis as a result of underlying IR.

  3. Direct Composite Laminate Veneers: Three Case Reports

    PubMed Central

    Korkut, Bora; Yanıkoğlu, Funda; Günday, Mahir

    2013-01-01

    Re-establishing a patient’s lost dental esthetic appearance is one of the most important topics for contemporary dentistry. New treatment materials and methods have been coming on the scene, day by day, in order to achieve such an aim. Most dentists prefer more conservative and aesthetic approaches, such as direct and indirect laminate veneer restorations, instead of full-ceramic crowns for anteriors where aesthetics is really important. Laminate veneers are restorations which are envisioned to correct existing abnormalities, esthetic deficiencies and discolo-rations. Laminate veneer restorations may be processed in two different ways: direct or indirect. Direct laminate veneers have no need to be prepared in the laboratory and are based on the principle of application of a composite material directly to the prepared tooth surface in the dental clinic. Indirect laminate veneers may be produced from composite materials or ceramics, which are cemented to the tooth with an adhesive resin. In this case report, direct composite laminate veneer technique used for three patients with esthetic problems related to fractures, discolorations and an old prolapsed restoration, is described and six-month follow-ups are discussed. As a conclusion, direct laminate veneer restorations may be a treatment option for patients with the esthetic problems of anterior teeth in cases similar to those reported here. PMID:23875090

  4. SENP1-modulated sumoylation regulates retinoblastoma protein (RB) and Lamin A/C interaction and stabilization.

    PubMed

    Sharma, P; Kuehn, M R

    2016-12-15

    The retinoblastoma tumor suppressor protein (RB) plays a critical role in cell proliferation and differentiation and its inactivation is a frequent underlying factor in tumorigenesis. While the regulation of RB function by phosphorylation is well studied, proteasome-mediated RB protein degradation is emerging as an important regulatory mechanism. Although our understanding of RB turnover is currently limited, there is evidence that the nuclear lamina filament protein Lamin A/C protects RB from proteasomal degradation. Here we show that SUMO1 conjugation of RB and Lamin A/C is modulated by the SUMO protease SENP1 and that sumoylation of both proteins is required for their interaction. Importantly, this SUMO1-dependent complex protects both RB and Lamin A/C from proteasomal turnover.

  5. Structural feasibility of parallel-laminated veneer crossarms

    Treesearch

    John Youngquist; Frank Brey; Joseph Jung

    1977-01-01

    Experimentally and commercially produced laminated M-19 crossarms were tested by standard Rural Electrification Administration (REA) crossarm tests. The laminated crossarms, produced by laminating veneer and by laminating solid-sawn dimension stock, generally performed satisfactorily according to REA specified standards. Materials tested are described and results on...

  6. A role for the p34cdc2 kinase and phosphatases in the regulation of phosphorylation and disassembly of lamin B2 during the cell cycle.

    PubMed Central

    Lüscher, B; Brizuela, L; Beach, D; Eisenman, R N

    1991-01-01

    While the p34cdc2 kinase is considered to be a critical regulator of mitosis, its function has not yet been directly linked to one of the key events during the onset of mitosis: nuclear envelope breakdown. Here we show that a major structural protein of the nuclear envelope, lamin B2, is phosphorylated by p34cdc2. Results from two-dimensional phosphopeptide mapping experiments demonstrate that the p34cdc2-specific phosphopeptides represent both mitotic and interphase specific phosphorylations of lamin B2 and include the major interphase phosphorylation site. In mitotic cells we detected two distinct forms of lamin B2 which differ in electrophoretic mobility and in degree of phosphorylation. The phosphorylation pattern of lamin B2 generated in vitro by p34cdc2 was more closely related to the less phosphorylated mitotic lamin B2, suggesting that another kinase(s) in addition to p34cdc2 is involved in generating the mitotic phosphorylation pattern. In addition, we show that treatment of interphase cells with okadaic acid, a potent phosphatase inhibitor, leads to the acquisition of mitosis-specific phosphopeptides and can reversibly increase the detergent-solubility of lamin B2. However, the M-phase-like phosphorylation of lamin B2 in itself is not sufficient to induce its disassembly from the nuclear lamina suggesting that an additional event(s) besides phosphorylation is required. Images PMID:1849074

  7. Damage of hybrid composite laminates

    NASA Astrophysics Data System (ADS)

    Haery, Haleh A.; Kim, Ho Sung

    2013-08-01

    Hybrid laminates consisting of woven glass fabric/epoxy composite plies and woven carbon fabric/epoxy composite plies are studied for fatigue damage and residual strength. A theoretical framework based on the systems approach is proposed as a guide to deal with the complexity involving uncertainties and a large number of variables in the hybrid composite system. A relative damage sensitivity factor expression was developed for quantitative comparisons between non-hybrid and hybrid composites. Hypotheses derived from the theoretical framework were tested and verified. The first hypothesis was that the difference between two different sets of properties produces shear stress in interface between carbon fibre reinforced plastics (CRP) and glass fibre reinforced plastics (GRP), and eventually become a source for CRP/GRP interfacial delamination or longitudinal cracking. The second hypothesis was that inter-fibre bundle delamination occurs more severely to CRP sub-system than GRP sub-system.

  8. Method of laminating structural members

    NASA Technical Reports Server (NTRS)

    Heier, W. C. (Inventor)

    1974-01-01

    A laminate is obtained by providing a lightweight core material, such as a honeycombed plastic or metal, within the cavity defined by an annular mold cavity frame. Face sheets, which are to be bonded to the core material, are provided on opposite sides of the frame and extend over the frame, thus sealing the core material in the cavity. An adhesive is provided between the core material and the face sheets and the combined thickness of the core material and adhesive is a close fit within the opposed face sheets. A gas tight seal, such as an O-ring gasket, is provided between the frame and the face sheet members to form a gas tight cavity between the face sheet members and the frame. External heat and pressure are used to bond the face sheets to the core material. Gas pressure is introduced into the sealed cavity to minimize out-gasing of the adhesive.

  9. Indentation law for composite laminates

    NASA Technical Reports Server (NTRS)

    Yang, S. H.

    1981-01-01

    Static indentation tests are described for glass/epoxy and graphite/epoxy composite laminates with steel balls as the indentor. Beam specimens clamped at various spans were used for the tests. Loading, unloading, and reloading data were obtained and fitted into power laws. Results show that: (1) contact behavior is not appreciably affected by the span; (2) loading and reloading curves seem to follow the 1.5 power law; and (3) unloading curves are described quite well by a 2.5 power law. In addition, values were determined for the critical indentation, alpha sub cr which can be used to predict permanent indentations in unloading. Since alpha sub cr only depends on composite material properties, only the loading and an unloading curve are needed to establish the complete loading-unloading-reloading behavior.

  10. Nuclear reactor safety device

    DOEpatents

    Hutter, E.

    1983-08-15

    A safety device is described for use in a nuclear reactor for axially repositioning a control rod with respect to the reactor core in the event of a thermal excursion. It comprises a laminated strip helically configured to form a tube, said tube being in operative relation to said control rod. The laminated strip is formed of at least two materials having different thermal coefficients of expansion, and is helically configured such that the material forming the outer lamina of the tube has a greater thermal coefficient of expansion than the material forming the inner lamina of said tube. In the event of a thermal excursion the laminated strip will tend to curl inwardly so that said tube will increase in length, whereby as said tube increases in length it exerts a force on said control rod to axially reposition said control rod with respect to said core.

  11. Nuclear reactor safety device

    DOEpatents

    Hutter, Ernest

    1986-01-01

    A safety device is disclosed for use in a nuclear reactor for axially repositioning a control rod with respect to the reactor core in the event of an upward thermal excursion. Such safety device comprises a laminated helical ribbon configured as a tube-like helical coil having contiguous helical turns with slidably abutting edges. The helical coil is disclosed as a portion of a drive member connected axially to the control rod. The laminated ribbon is formed of outer and inner laminae. The material of the outer lamina has a greater thermal coefficient of expansion than the material of the inner lamina. In the event of an upward thermal excursion, the laminated helical coil curls inwardly to a smaller diameter. Such inward curling causes the total length of the helical coil to increase by a substantial increment, so that the control rod is axially repositioned by a corresponding amount to reduce the power output of the reactor.

  12. Flat laminated microbial mat communities

    NASA Astrophysics Data System (ADS)

    Franks, Jonathan; Stolz, John F.

    2009-10-01

    Flat laminated microbial mats are complex microbial ecosystems that inhabit a wide range of environments (e.g., caves, iron springs, thermal springs and pools, salt marshes, hypersaline ponds and lagoons, methane and petroleum seeps, sea mounts, deep sea vents, arctic dry valleys). Their community structure is defined by physical (e.g., light quantity and quality, temperature, density and pressure) and chemical (e.g., oxygen, oxidation/reduction potential, salinity, pH, available electron acceptors and donors, chemical species) parameters as well as species interactions. The main primary producers may be photoautotrophs (e.g., cyanobacteria, purple phototrophs, green phototrophs) or chemolithoautophs (e.g., colorless sulfur oxidizing bacteria). Anaerobic phototrophy may predominate in organic rich environments that support high rates of respiration. These communities are dynamic systems exhibiting both spatial and temporal heterogeneity. They are characterized by steep gradients with microenvironments on the submillimeter scale. Diel oscillations in the physical-chemical profile (e.g., oxygen, hydrogen sulfide, pH) and species distribution are typical for phototroph-dominated communities. Flat laminated microbial mats are often sites of robust biogeochemical cycling. In addition to well-established modes of metabolism for phototrophy (oxygenic and non-oxygenic), respiration (both aerobic and anaerobic), and fermentation, novel energetic pathways have been discovered (e.g., nitrate reduction couple to the oxidation of ammonia, sulfur, or arsenite). The application of culture-independent techniques (e.g., 16S rRNA clonal libraries, metagenomics), continue to expand our understanding of species composition and metabolic functions of these complex ecosystems.

  13. Neuropathic pain management in chronic laminitis.

    PubMed

    Driessen, Bernd; Bauquier, Sébastien H; Zarucco, Laura

    2010-08-01

    Managing pain in horses afflicted by chronic laminitis is one of the greatest challenges in equine clinical practice because it is the dreadful suffering of the animals that most often forces the veterinarian to end the battle with this disease. The purpose of this review is to summarize our current understanding of the complex mechanisms involved in generating and amplifying pain in animals with laminitis and, based on this information, to propose a modified approach to pain therapy. Furthermore, a recently developed pain scoring technique is presented that may help better quantify pain and the monitoring of responses to analgesic treatment in horses with laminitis.

  14. Thermal buckling of symmetrically laminated composite plates

    NASA Technical Reports Server (NTRS)

    Meyers, C. A.; Hyer, M. W.

    1991-01-01

    This paper discusses an investigation into the thermal buckling of symmetrically laminated composite plates. In this study thermal buckling is investigated for laminates under two different simple support conditions, fixed and sliding. These laminates are subjected to the conditions of a uniform temperature change, and a temperature change varying linearly along the length of the plate. The effects of the principal material axes not being aligned with the edges of the plate are also investigated. The buckling response is studied using variational methods, specifically the Trefftz criterion. A Rayleigh-Ritz formulation is used to obtain numerical results from the formulations for the prebuckling response and the buckling response.

  15. Thermal buckling of laminated composite shells

    SciTech Connect

    Thangaratnam, R.K.; Palaninathan, R.; Ramachandran, J. )

    1990-05-01

    The linear buckling analysis of laminated composite cylindrical and conical shells under thermal load using the finite element method is reported here. Critical temperatures are presented for various cases of cross-ply and angly-ply laminated shells. The effects of radius/thickness ratio, number of layers, ratio of coefficients of thermal expansion, and the angle of fiber orientation have been studied. The results indicate that the buckling behavior of laminated shell under thermal load is different from that of mechanically loaded shell with respect to the angle of fiber orientation. 6 refs.

  16. NOLIN: A nonlinear laminate analysis program

    NASA Technical Reports Server (NTRS)

    Kibler, J. J.

    1975-01-01

    A nonlinear, plane-stress, laminate analysis program, NOLIN, was developed which accounts for laminae nonlinearity under inplane shear and transverse extensional stress. The program determines the nonlinear stress-strain behavior of symmetric laminates subjected to any combination of inplane shear and biaxial extensional loadings. The program has the ability to treat different stress-strain behavior in tension and compression, and predicts laminate failure using any or all of maximum stress, maximum strain, and quadratic interaction failure criteria. A brief description of the program is presented including discussion of the flow of information and details of the input required. Sample problems and a complete listing of the program is also provided.

  17. Decreased Bone Formation and Osteopenia in Lamin A/C-Deficient Mice

    PubMed Central

    Vidal, Christopher; McCorquodale, Thomas; Herrmann, Markus; Fatkin, Diane; Duque, Gustavo

    2011-01-01

    Age-related bone loss is associated with changes in bone cellularity with characteristically low levels of osteoblastogenesis. The mechanisms that explain these changes remain unclear. Although recent in vitro evidence has suggested a new role for proteins of the nuclear envelope in osteoblastogenesis, the role of these proteins in bone cells differentiation and bone metabolism in vivo remains unknown. In this study, we used the lamin A/C null (Lmna−/−) mice to identify the role of lamin A/C in bone turnover and bone structure in vivo. At three weeks of age, histological and micro computed tomography measurements of femurs in Lmna−/− mice revealed a significant decrease in bone mass and microarchitecture in Lmna−/− mice as compared with their wild type littermates. Furthermore, quantification of cell numbers after normalization with bone surface revealed a significant reduction in osteoblast and osteocyte numbers in Lmna−/− mice compared with their WT littermates. In addition, Lmna−/− mice have significantly lower osteoclast number, which show aberrant changes in their shape and size. Finally, mechanistic analysis demonstrated that absence of lamin A/C is associated with increase expression of MAN-1 a protein of the nuclear envelope closely regulated by lamin A/C, which also colocalizes with Runx2 thus affecting its capacity as osteogenic transcription factor. In summary, these data clearly indicate that the presence of lamin A/C is necessary for normal bone turnover in vivo and that absence of lamin A/C induces low bone turnover osteopenia resembling the cellular changes of age-related bone loss. PMID:21547077

  18. Home care for horses with chronic laminitis.

    PubMed

    Orsini, James A; Wrigley, Jennifer; Riley, Patrick

    2010-04-01

    Home care for horses with chronic laminitis has been discussed rarely in the veterinary literature even though, at any given time, most of us have at least 1 chronic laminitis case in our care that is being managed at home by the owner. Almost all of our knowledge on this aspect of laminitis treatment has been gleaned through experience, by individually working through the medical, ethical, financial, and emotional challenges these cases can present. Much has already been presented on the medical management of the laminitic horse and on strategies for trimming and shoeing the laminitic foot. This article focuses on the other challenges so often faced when directing the home care of a horse with chronic laminitis.

  19. Pattern recognition of laminated sediments methodology

    NASA Astrophysics Data System (ADS)

    Barba-Rojo, Perla Karina; Solorza-Calderón, Selene; González-Fernández, Antonio

    2016-12-01

    This work presents a different aproach for laminae counting and thickness measurements on laminated sediment images. This is done by the use of morphological operations and minimum variance quantization.

  20. Free edge effects in laminated composites

    NASA Technical Reports Server (NTRS)

    Herakovich, C. T.

    1989-01-01

    The fundamental mechanics of free-edge effects in laminated fiber-reinforced composites is examined, reviewing the results of recent experimental and analytical investigations. The derivation of the governing equations for the basic problem is outlined, including the equilibrium and mismatch conditions and the elasticity formulation, and experimental data on axial displacement and shear strain in angle-ply laminates are summarized. Numerical predictions of free-edge deformation and interlaminar and through-thickness stress distributions are presented for cross-ply, angle-ply, and quasi-isotropic laminates, and the mechanisms of edge damage and failure in angle-ply laminates are briefly characterized. Extensive diagrams, drawings, graphs, and photographs are provided.

  1. Method for fabricating laminated uranium composites

    DOEpatents

    Chapman, L.R.

    1983-08-03

    The present invention is directed to a process for fabricating laminated composites of uranium or uranium alloys and at least one other metal or alloy. The laminated composites are fabricated by forming a casting of the molten uranium with the other metal or alloy which is selectively positioned in the casting and then hot-rolling the casting into a laminated plate in or around which the casting components are metallurgically bonded to one another to form the composite. The process of the present invention provides strong metallurgical bonds between the laminate components primarily since the bond disrupting surface oxides on the uranium or uranium alloy float to the surface of the casting to effectively remove the oxides from the bonding surfaces of the components.

  2. TRF2 and lamin A/C interact to facilitate the functional organization of chromosome ends

    PubMed Central

    Wood, Ashley M.; Danielsen, Jannie M. Rendtlew; Lucas, Catherine A.; Rice, Ellen L.; Scalzo, David; Shimi, Takeshi; Goldman, Robert D.; Smith, Erica D.; Le Beau, Michelle M.; Kosak, Steven T.

    2014-01-01

    Telomeres protect the ends of linear genomes, and the gradual loss of telomeres is associated with cellular ageing. Telomere protection involves the insertion of the 3′ overhang facilitated by telomere repeat-binding factor 2 (TRF2) into telomeric DNA, forming t-loops. We present evidence suggesting that t-loops can also form at interstitial telomeric sequences in a TRF2-dependent manner, forming an interstitial t-loop (ITL). We demonstrate that TRF2 association with interstitial telomeric sequences is stabilized by co-localization with A-type lamins (lamin A/C). We also find that lamin A/C interacts with TRF2 and that reduction in levels of lamin A/C or mutations in LMNA that cause an autosomal dominant premature ageing disorder—Hutchinson Gilford Progeria Syndrome (HGPS)—lead to reduced ITL formation and telomere loss. We propose that cellular and organismal ageing are intertwined through the effects of the interaction between TRF2 and lamin A/C on chromosome structure. PMID:25399868

  3. Fatigue crack growth in aluminum laminate composites

    SciTech Connect

    Hoffman, P.B.; Carpenter, R.D.; Gibeling, J.C.

    1996-12-31

    Fatigue crack growth has been measured in a laminated metal composite (LMC) consisting of alternating layers of AA6090/SiC/25p metal matrix composite (MMC) and AA5182 alloy. This material was tested in both as-pressed (F temper) and aged (T6 temper) conditions. Corresponding crack growth measurements were made in self-laminates of both the MMC and AA5182 materials to examine the role of the interfaces.

  4. Descriptive epidemiological study of equine laminitis.

    PubMed

    Slater, M R; Hood, D M; Carter, G K

    1995-09-01

    A descriptive and matched case-control study of laminitis was conducted in 7 private practices and at the Texas Veterinary Medical Centre (TVMC) between May 1992 and July 1993. Out of 108 horses with laminitis, 19 acute (49%) and 20 chronic (51%) cases were seen in private practice and 16 acute (23%) and 53 (77%) cases at the TVMC. Gastrointestinal disease was the most common problem in 19/35 horses (54%), occurring just prior to the onset of acute laminitis in all hospitals. Among all horses in the study, most commonly used drugs were phenylbutazone (68%), acepromazine (34%), dimethyl sulphoxide (DMSO) (27%), antibiotics of various types (19%) and flunixin meglumine (19%). Acepromazine, DMSO and flunixin meglumine were used more commonly in acute cases of laminitis compared to chronic cases. In acutely affected horses, DMSO and flunixin meglumine were used significantly more often at the TVMC. In chronic cases, phenylbutazone and antibiotics were used more often in private practice. Shoeing and trimming were more commonly part of the treatment protocol for chronic cases. There were no significant associations between age, breed, sex or weight and the occurrence of acute laminitis. Horses with chronic laminitis were significantly older (P=0.04) and more females tended to be affected (P=0.08).

  5. Phosphorylation statuses at different residues of lamin B2, B1, and A/C dynamically and independently change throughout the cell cycle

    SciTech Connect

    Kuga, Takahisa; Nozaki, Naohito; Matsushita, Kazuyuki; Nomura, Fumio; Tomonaga, Takeshi

    2010-08-15

    Lamins, major components of the nuclear lamina, undergo phosphorylation at multiple residues during cell cycle progression, but their detailed phosphorylation kinetics remain largely undetermined. Here, we examined changes in the phosphorylation of major phosphorylation residues (Thr14, Ser17, Ser385, Ser387, and Ser401) of lamin B2 and the homologous residues of lamin B1, A/C during the cell cycle using novel antibodies to the site-specific phosphorylation. The phosphorylation levels of these residues independently changed during the cell cycle. Thr14 and Ser17 were phosphorylated during G{sub 2}/M phase to anaphase/telophase. Ser385 was persistently phosphorylated during mitosis to G{sub 1} phase, whereas Ser387 was phosphorylated discontinuously in prophase and G{sub 1} phase. Ser401 phosphorylation was enhanced in the G{sub 1}/S boundary. Immunoprecipitation using the phospho-antibodies suggested that metaphase-phosphorylation at Thr14, Ser17, and Ser385 of lamins occurred simultaneously, whereas G{sub 1}-phase phosphorylation at Ser385 and Ser387 occurred in distinct pools or with different timings. Additionally, we showed that lamin B2 phosphorylated at Ser17, but not Ser385, Ser387 and Ser401, was exclusively non-ionic detergent soluble, depolymerized forms in growing cells, implicating specific involvement of Ser17 phosphorylation in lamin depolymerization and nuclear envelope breakdown. These results suggest that the phosphorylations at different residues of lamins might play specific roles throughout the cell cycle.

  6. Bovine laminitis: clinical aspects, pathology and pathogenesis with reference to acute equine laminitis.

    PubMed

    Boosman, R; Németh, F; Gruys, E

    1991-07-01

    This review deals with the features of clinical and subclinical laminitis in cattle. Prominent clinical signs of acute laminitis are a tender gait and arched back. The sole horn reveals red and yellowish discolourations within five days. In subacute and chronic cases clinical signs are less severe. In chronic laminitis the shape of the claws is altered. Laminitis is frequently followed by sole ulceration and white zone lesions. Blood tests showed no significant changes for laminitic animals. Arteriographic studies of claws affected by laminitis indicated that blood vessels had narrowed lumens. Gross pathology revealed congestion of the corium and rotation of the distal phalanx. Histopathologic studies indicate that laminitis is associated with changes of the vasculature. Peripartum management and nutrition are important factors in its aetiology. It is hypothesised that laminitis is evoked by disturbed digital circulation. In the pathogenesis of acute laminitis three factors are considered important: the occurrence of thrombosis, haemodynamic aspects of the corium, and endotoxins which trigger these pathologic events.

  7. Effect of laminate edge conditions on the formation of microvoids in composite laminates

    NASA Astrophysics Data System (ADS)

    Anderson, J. P.; Altan, M. C.

    2015-05-01

    Manufacturing defects such as microvoids are common in thermoset composite components and are known to negatively affect their strength. The resin pressure developed in and the resin flow out from the laminates during cure have been reported to be the primary factors influencing the final void content of a composite component. In this work, the effect of laminate edge conditions during the cure process on the formation of microvoids was experimentally investigated. This was achieved by fabricating eight-ply laminates from TenCate® BT250/7781 prepreg in a hot-press at a constant cure pressure of 170 kPa while limiting the laminate perimeter available for resin flow by 0%, 25%, 50%, 75%, and 100%. The individual plies of these five laminates were conditioned at 99% relative humidity before curing to maximize the moisture present in the lay-up before fabrication. The presence of moisture in the lay-ups was expected to promote void formation and allow the effect of restricting flow at the edges of a laminate to be better identified. The restriction of resin outflow was found to cause the average characteristic void diameter to decrease by 17% and void content to rise by 33%. This phenomenon was identified to be a result of the outflow restriction increasing the number of voids trapped within the laminate and indicates that for laminates cured at low pressures resin outflow is the dominant mechanism for void reduction.

  8. Distinct Fiber Type Signature in Mouse Muscles Expressing a Mutant Lamin A Responsible for Congenital Muscular Dystrophy in a Patient.

    PubMed

    Barateau, Alice; Vadrot, Nathalie; Agbulut, Onnik; Vicart, Patrick; Batonnet-Pichon, Sabrina; Buendia, Brigitte

    2017-04-24

    Specific mutations in LMNA, which encodes nuclear intermediate filament proteins lamins A/C, affect skeletal muscle tissues. Early-onset LMNA myopathies reveal different alterations of muscle fibers, including fiber type disproportion or prominent dystrophic and/or inflammatory changes. Recently, we identified the p.R388P LMNA mutation as responsible for congenital muscular dystrophy (L-CMD) and lipodystrophy. Here, we asked whether viral-mediated expression of mutant lamin A in murine skeletal muscles would be a pertinent model to reveal specific muscle alterations. We found that the total amount and size of muscle fibers as well as the extent of either inflammation or muscle regeneration were similar to wildtype or mutant lamin A. In contrast, the amount of fast oxidative muscle fibers containing myosin heavy chain IIA was lower upon expression of mutant lamin A, in correlation with lower expression of genes encoding transcription factors MEF2C and MyoD. These data validate this in vivo model for highlighting distinct muscle phenotypes associated with different lamin contexts. Additionally, the data suggest that alteration of muscle fiber type identity may contribute to the mechanisms underlying physiopathology of L-CMD related to R388P mutant lamin A.

  9. Treating laminitis: beyond the mechanics of trimming and shoeing.

    PubMed

    Baker, William R

    2012-08-01

    Laminitis is typically classified into developmental or prodromal, acute, subacute, and chronic phases. Scientific evidence regarding the pathophysiology of laminitis does exist, but it is often conflicting and dependent on the clinician's interpretation/understanding of the study or the model used for inducing laminitis. The diagnosis of laminitis consists of obtaining an accurate history, performing a thorough physical examination, and taking good-quality radiographs. The use of radiographs for diagnosis and interpretation of laminitis is an absolute necessity for the clinician. Laminitis is one disease that requires the assembly of a team consisting of the veterinarian, the farrier, and the owner to be successfully treated.

  10. Finite element analysis of shear deformable laminated composite plates

    SciTech Connect

    Kam, T.Y.; Chang, R.R. )

    1993-03-01

    A shear deformable finite element is developed for the analysis of thick laminated composite plates. The finite element formulation is based on Mindlin's plate theory in which shear correction factors are derived from the exact expressions for orthotropic materials. The element is used to solve a variety of problems on deflection, stress distribution, natural frequency and buckling of laminated composite plates. The effects of material properties, plate aspect ratio, length-to-thickness ratio, number of layers and lamination angle on the mechanical behaviors of laminated composite plates are investigated. Optimal lamination arrangements of layers for laminated composite plates of particular applications are determined.

  11. Local Translation of Extranuclear Lamin B Promotes Axon Maintenance

    PubMed Central

    Yoon, Byung C.; Jung, Hosung; Dwivedy, Asha; O'Hare, Catherine M.; Zivraj, Krishna H.; Holt, Christine E.

    2012-01-01

    Summary Local protein synthesis plays a key role in regulating stimulus-induced responses in dendrites and axons. Recent genome-wide studies have revealed that thousands of different transcripts reside in these distal neuronal compartments, but identifying those with functionally significant roles presents a challenge. We performed an unbiased screen to look for stimulus-induced, protein synthesis-dependent changes in the proteome ofXenopus retinal ganglion cell (RGC) axons. The intermediate filament protein lamin B2 (LB2), normally associated with the nuclear membrane, was identified as an unexpected major target. Axonal ribosome immunoprecipitation confirmed translation of lb2 mRNA in vivo. Inhibition of lb2 mRNA translation in axons in vivo does not affect guidance but causes axonal degeneration. Axonal LB2 associates with mitochondria, and LB2-deficient axons exhibit mitochondrial dysfunction and defects in axonal transport. Our results thus suggest that axonally synthesized lamin B plays a crucial role in axon maintenance by promoting mitochondrial function. PMID:22341447

  12. Regulation of prelamin A but not lamin C by miR-9, a brain-specific microRNA.

    PubMed

    Jung, Hea-Jin; Coffinier, Catherine; Choe, Youngshik; Beigneux, Anne P; Davies, Brandon S J; Yang, Shao H; Barnes, Richard H; Hong, Janet; Sun, Tao; Pleasure, Samuel J; Young, Stephen G; Fong, Loren G

    2012-02-14

    Lamins A and C, alternatively spliced products of the LMNA gene, are key components of the nuclear lamina. The two isoforms are found in similar amounts in most tissues, but we observed an unexpected pattern of expression in the brain. Western blot and immunohistochemistry studies showed that lamin C is abundant in the mouse brain, whereas lamin A and its precursor prelamin A are restricted to endothelial cells and meningeal cells and are absent in neurons and glia. Prelamin A transcript levels were low in the brain, but this finding could not be explained by alternative splicing. In lamin A-only knockin mice, where alternative splicing is absent and all the output of the gene is channeled into prelamin A transcripts, large amounts of lamin A were found in peripheral tissues, but there was very little lamin A in the brain. Also, in knockin mice expressing exclusively progerin (a toxic form of prelamin A found in Hutchinson-Gilford progeria syndrome), the levels of progerin in the brain were extremely low. Further studies showed that prelamin A expression, but not lamin C expression, is down-regulated by a brain-specific microRNA, miR-9. Expression of miR-9 in cultured cells reduced lamin A expression, and this effect was abolished when the miR-9-binding site in the prelamin A 3' UTR was mutated. The down-regulation of prelamin A expression in the brain could explain why mouse models of Hutchinson-Gilford progeria syndrome are free of central nervous system pathology.

  13. Lamin A/C Acts as an Essential Factor in Mesenchymal Stem Cell Differentiation Through the Regulation of the Dynamics of the Wnt/β-Catenin Pathway.

    PubMed

    Bermeo, Sandra; Vidal, Christopher; Zhou, Hong; Duque, Gustavo

    2015-10-01

    Changes in the expression of lamin A/C, a fibrilar protein of the nuclear envelope, are associated with the cellular features of age-related bone loss. Reduced expression of lamin A/C inhibits osteoblastogenesis while facilitating adipogenic differentiation of mesenchymal stem cells (MSC) in vitro and in vivo. In this study we investigated the regulatory role that lamin A/C plays on the essential elements of the Wnt/β-catenin pathway, which are pivotal in MSC differentiation. Initially, we assessed the effect of lamin A/C gene (LMNA) overexpression on MSC differentiation while compared it to lamin A/C depleted MSC. Osteogenesis and gene expression of osteogenic factors were higher in LMNA-transfected MSC as compared to control. Conversely, adipogenesis and expression of adipogenic factors were significantly lower in LMNA transfected cells. Nuclear β-catenin was significantly higher (∼two fold) in MSC expressing higher levels of LMNA as compared to control with nuclear β-catenin levels being significantly lower (∼ -42%) in siRNA-treated MSC. Luciferase activity for β-catenin-mediated transcriptional activation was significantly higher in cells overexpressing LMNA. These data indicate that MSC overexpressing LMNA have higher osteogenic and lower adipogenic differentiation potential. In conclusion, our studies demonstrate that lamin A/C plays a significant role in the differentiation of both osteoblasts and adipocytes by regulating some of the elements of Wnt/β-catenin signaling during early MSC differentiation.

  14. Mechanical Behavior of Fabric-Film Laminates

    NASA Technical Reports Server (NTRS)

    Said, Magdi S.

    1999-01-01

    Inflatable structures are gaining wide support in planetary scientific missions as well as commercial applications. For such applications a new class of materials made of laminating thin homogenous films to lightweight fabrics are being considered us structura1 gas envelops. The emerging composite materials are a result of recent advances in the manufacturing cf 1ightweight, high strength fibers, fabrics and scrims. The lamination of these load-carrying members with the proper gas barrier film results in wide range of materials suitable for various loading and environmental conditions. Polyester - based woven fabrics laminated to thin homogeneus film of polyester (Maylar) is an example of this class. This fabric/ film laminate is being considered for the development a material suitable for building large gas envelopes for use in the NASA Ultra Long Duration Balloon Program (ULDB). Compared to commercial homogeneus films, the material provides relatively high strength to weight ratio as well as better resistance to crack and tear propagation. The purpose of this papers is to introduce the mechanical behavior of this class of multi-layers composite and to highlight some of the concerns observed during the characterization of these laminate composites.

  15. Fracture behavior of hybrid composite laminates

    NASA Technical Reports Server (NTRS)

    Kennedy, J. M.

    1983-01-01

    The tensile fracture behavior of 15 center-notched hybrid laminates was studied. Three basic laminate groups were tested: (1) a baseline group with graphite/epoxy plies, (2) a group with the same stacking sequence but where the zero-deg plies were one or two plies of S-glass or Kevlar, and (3) a group with graphite plies but where the zero-deg plies were sandwiched between layers of perforated Mylar. Specimens were loaded linearly with time; load, far field strain, and crack opening displacement (COD) were monitored. The loading was stopped periodically and the notched region was radiographed to reveal the extent and type of damage (failure progression). Results of the tests showed that the hybrid laminates had higher fracture toughnesses than comparable all-graphite laminates. The higher fracture toughness was due primarily to the larger damage region at the ends of the slit; delamination and splitting lowered the stress concentration in the primary load-carrying plies. A linear elastic fracture analysis, which ignored delamination and splitting, underestimated the fracture toughness. For almost all of the laminates, the tests showed that the fracture toughness increased with crack length. The size of the damage region at the ends of the slit and COD measurements also increased with crack length.

  16. Lamination residual stresses in fiber composites

    NASA Technical Reports Server (NTRS)

    Daniel, I. M.; Liber, T.

    1975-01-01

    An experimental investigation was conducted to determine the magnitude of lamination residual stresses in angle-ply composites and to evaluate their effects on composite structural integrity. The materials investigated were boron/epoxy, boron/polyimide, graphite/low modulus epoxy, graphite/high modulus epoxy, graphite/polyimide and s-glass/epoxy. These materials were fully characterized. Static properties of laminates were also determined. Experimental techniques using embedded strain gages were developed and used to measure residual strains during curing. The extent of relaxation of lamination residual stresses was investigated. It was concluded that the degree of such relaxation is low. The behavior of angle-ply laminates subjected to thermal cycling, tensile load cycling, and combined thermal cycling with tensile load was investigated. In most cases these cycling programs did not have any measurable influence on residual strength and stiffness of the laminates. In the tensile load cycling tests, the graphite/polyimide shows the highest endurance with 10 million cycle runouts at loads up to 90 percent of the static strength.

  17. Evaluation of Behaviours of Laminated Glass

    NASA Astrophysics Data System (ADS)

    Sable, L.; Japins, G.; Kalnins, K.

    2015-11-01

    Visual appearance of building facades and other load bearing structures, which now are part of modern architecture, is the reason why it is important to investigate in more detail the reliability of laminated glass for civil structures. Laminated glass in particular has become one of the trendy materials, for example Apple© stores have both load carrying capacity and transparent appearance. Glass has high mechanical strength and relatively medium density, however, the risk of sudden brittle failure like concrete or other ceramics determine relatively high conservatism in design practice of glass structures. This should be changed as consumer requirements evolve calling for a safe and reliable design methodology and corresponding building standards. A design methodology for glass and glass laminates should be urgently developed and included as a chapter in Eurocode. This paper presents initial experimental investigation of behaviour of simple glass sheets and laminated glass samples in 4-point bending test. The aim of the current research is to investigate laminated glass characteristic values and to verify the obtained experimental results with finite element method for glass and EVA material in line with future European Structural Design of Glass Components code.

  18. Down-regulation of the Lamin A/C in neuroblastoma triggers the expansion of tumor initiating cells.

    PubMed

    Nardella, Marta; Guglielmi, Loredana; Musa, Carla; Iannetti, Ilaria; Maresca, Giovanna; Amendola, Donatella; Porru, Manuela; Carico, Elisabetta; Sessa, Giuseppe; Camerlingo, Rosalba; Dominici, Carlo; Megiorni, Francesca; Milan, Marika; Bearzi, Claudia; Rizzi, Roberto; Pirozzi, Giuseppe; Leonetti, Carlo; Bucci, Barbara; Mercanti, Delio; Felsani, Armando; D'Agnano, Igea

    2015-10-20

    Tumor-initiating cells constitute a population within a tumor mass that shares properties with normal stem cells and is considered responsible for therapy failure in many cancers. We have previously demonstrated that knockdown of the nuclear envelope component Lamin A/C in human neuroblastoma cells inhibits retinoic acid-mediated differentiation and results in a more aggressive phenotype. In addition, Lamin A/C is often lost in advanced tumors and changes in the nuclear envelope composition occur during tumor progression. Based on our previous data and considering that Lamin A/C is expressed in differentiated tissues, we hypothesize that the lack of Lamin A/C could predispose cells toward a stem-like phenotype, thus influencing the development of tumor-initiating cells in neuroblastoma. This paper demonstrates that knockdown of Lamin A/C triggers the development of a tumor-initiating cell population with self-renewing features in human neuroblastoma cells. We also demonstrates that the development of TICs is due to an increased expression of MYCN gene and that in neuroblastoma exists an inverse relationship between LMNA and MYCN expression.

  19. Silicone modified resins for graphite fiber laminates

    NASA Technical Reports Server (NTRS)

    Frost, L. W.; Bower, G. M.

    1979-01-01

    The development of silicon modified resins for graphite fiber laminates which will prevent the dispersal of graphite fibers when the composites are burned is discussed. Eighty-five silicone modified resins were synthesized and evaluated including unsaturated polyesters, thermosetting methacrylates, epoxies, polyimides, and phenolics. Neat resins were judged in terms of Si content, homogeneity, hardness, Char formation, and thermal stability. Char formation was estimated by thermogravimetry to 1,000 C in air and in N2. Thermal stability was evaluated by isothermal weight loss measurements for 200 hrs in air at three temperatures. Four silicone modified epoxies were selected for evaluation in unidirectional filament wound graphite laminates. Neat samples of these resins had 1,000 C char residues of 25 to 50%. The highest flexural values measured for the laminates were a strength of 140 kpsi and a modulus of 10 Mpsi. The highest interlaminar shear strength was 5.3 kpsi.

  20. Damage initiation and propagation in metal laminates

    SciTech Connect

    Riddle, R.A.; Lesuer, D.R.; Syn, C.K.

    1996-07-26

    The metal laminates proposed here for aircraft structures are Al alloy interlayers between Al alloy based metal matrix composite (MMC) plates reinforced with Si carbide particles. Properties to be tailored for jet engine fan containment and wing and auxiliary support structures include the important property fracture toughness. A method was developed for simulating and predicting crack initiation/growth using finite element analysis and fracture mechanics. An important key in predicting the failure is the tie- break slideline with prescribed (chosen based on J Integral calculations) effective plastic strain to failure in elements along the slideline. More development of the method is needed, particularly in its correlation with experimental data from various fracture toughness and strength tests of metal laminates. Results show that delamination at the interface of the ductile interlayer and MMC material can add significantly to the energy required to propagate a crack through a metal laminate. 11 figs, 7 refs.

  1. Pasture nonstructural carbohydrates and equine laminitis.

    PubMed

    Longland, Annette C; Byrd, Bridgett M

    2006-07-01

    Fresh forages constitute a majority of the diet for many horses and ponies that graze on pastures during the growing season in many parts of the world. Grasses generally predominate in such pastures, with varying proportions of legumes. Nonstructural carbohydrates (NSC) (simple sugars, starch, and fructan) can induce laminitis experimentally, and NSC can accumulate to >400 g/kg of dry matter (DM) in pasture grasses. In this article we discuss the environmental factors affecting NSC accumulation in pastures and estimate the potential daily intakes of pasture NSC by grazing horses. We also discuss strategies for both reducing the NSC content of pastures and management practices that can help reduce intakes of pasture NSC by equines at risk of developing laminitis. This study reveals the importance of accurate forage analysis in the development of feeding regimens for equines at risk of laminitis.

  2. Flutter of laminated plates in supersonic flow

    NASA Technical Reports Server (NTRS)

    Sawyer, J. W.

    1975-01-01

    A solution procedure was developed using linear small deflection theory for the flutter of simply supported laminated plates. For such plates, the bending and extensional governing equations are coupled and have cross-stiffness terms which do not appear in classical plate theory. An extended Galerkin method is used to obtain approximate solutions to the governing equations, and the aerodynamic pressure loading used in the analysis is that given by linear piston theory with flow at arbitrary cross-flow angle. A limited parametric study was conducted for typical laminated composite plates. The calculations show that both the bending-extensional coupling and the cross-stiffness terms have a large destabilizing effect on flutter. Since classical plate theory does not consider bending-extensional coupling and cross stiffness terms, it usually gives inaccurate and nonconservative flutter boundaries for laminated plates.

  3. Hybrid Laminates for Application in North Conditions

    NASA Astrophysics Data System (ADS)

    Antipov, V. V.; Oreshko, E. I.; Erasov, V. S.; Serebrennikova, N. Yu.

    2016-11-01

    A hybrid aluminum-lithium alloy/SIAL laminate as a possible material for application in structures operated in North conditions is considered. The finite-element method is used for a buckling stability analysis of hybrid panels, bars, and plates. A technique allowing one to compare the buckling stability of multilayered hybrid plates is offered. Compression tests were run on a hybrid laminate wing panel as a prototype of the top panel of TU-204SM airplane made from a high-strength B95T2 aluminum alloy. It turned out that the lighter composite panel had a higher load-carrying capacity than the aluminum one. Results of investigation into the properties the hybrid aluminum-lithium alloy/SIAL laminate and an analysis of scientific-technical data on this subject showed that this composite material could be used in the elements of airframes, including those operated in north conditions.

  4. Plated lamination structures for integrated magnetic devices

    SciTech Connect

    Webb, Bucknell C.

    2014-06-17

    Semiconductor integrated magnetic devices such as inductors, transformers, etc., having laminated magnetic-insulator stack structures are provided, wherein the laminated magnetic-insulator stack structures are formed using electroplating techniques. For example, an integrated laminated magnetic device includes a multilayer stack structure having alternating magnetic and insulating layers formed on a substrate, wherein each magnetic layer in the multilayer stack structure is separated from another magnetic layer in the multilayer stack structure by an insulating layer, and a local shorting structure to electrically connect each magnetic layer in the multilayer stack structure to an underlying magnetic layer in the multilayer stack structure to facilitate electroplating of the magnetic layers using an underlying conductive layer (magnetic or seed layer) in the stack as an electrical cathode/anode for each electroplated magnetic layer in the stack structure.

  5. Development of tough, moisture resistant laminating resins

    NASA Technical Reports Server (NTRS)

    Brand, R. A.; Harrison, E. S.

    1982-01-01

    Tough, moisture resistant laminating resins for employment with graphite fibers were developed. The new laminating resins exhibited cost, handleability and processing characteristics equivalent to 394K (250 F) curing epoxies. The laminating resins were based on bisphenol A dicyanate and monofunctional cyanates with hydrophobic substituents. These resins sorb only small quantities of moisture at equilibrium (0.5% or less) with minimal glass transition temperature depression and represent an improvement over epoxies which sorb around 2% moisture at equilibrium. Toughening was accomplished by the precipitation of small diameter particles of butadiene nitrile rubber throughout the resin matrix. The rubber domains act as microcrack termini and energy dissipation sites, allowing increased stress accommodation prior to catastrophic failure. A unique blend of amine terminated butadiene nitrile elastomer (MW 2,000) and a high nitrile content butadiene nitrile rubber yielded the desired resin morphology.

  6. Preliminary evaluation of hybrid titanium composite laminates

    NASA Technical Reports Server (NTRS)

    Miller, J. L.; Progar, D. J.; Johnson, W. S.; St.clair, T. L.

    1994-01-01

    In this study, the mechanical response of hybrid titanium composite laminates (HTCL) was evaluated at room and elevated temperatures. Also, the use of an elastic-plastic laminate analysis program for predicting the tensile response from constituent properties was verified. The improvement in mechanical properties achieved by the laminates was assessed by comparing the results of static strength and constant amplitude fatigue tests to those for monolithic titanium sheet. Two HTCL were fabricated with different fiber volume fractions, resin layer thicknesses, and resins. One panel was thicker and was more poorly bonded in comparison to other. Consequently, the former had a lower tensile strength, while fewer cracks grew in this panel and at a slower rate. Both panels showed an improvement in fatigue life of almost two orders of magnitude. The model predictions were also in good agreement with the experimental results for both HTCL panels.

  7. Structural reliability analysis of laminated CMC components

    NASA Technical Reports Server (NTRS)

    Duffy, Stephen F.; Palko, Joseph L.; Gyekenyesi, John P.

    1991-01-01

    For laminated ceramic matrix composite (CMC) materials to realize their full potential in aerospace applications, design methods and protocols are a necessity. The time independent failure response of these materials is focussed on and a reliability analysis is presented associated with the initiation of matrix cracking. A public domain computer algorithm is highlighted that was coupled with the laminate analysis of a finite element code and which serves as a design aid to analyze structural components made from laminated CMC materials. Issues relevant to the effect of the size of the component are discussed, and a parameter estimation procedure is presented. The estimation procedure allows three parameters to be calculated from a failure population that has an underlying Weibull distribution.

  8. Stochastic damage evolution in textile laminates

    NASA Technical Reports Server (NTRS)

    Dzenis, Yuris A.; Bogdanovich, Alexander E.; Pastore, Christopher M.

    1993-01-01

    A probabilistic model utilizing random material characteristics to predict damage evolution in textile laminates is presented. Model is based on a division of each ply into two sublaminas consisting of cells. The probability of cell failure is calculated using stochastic function theory and maximal strain failure criterion. Three modes of failure, i.e. fiber breakage, matrix failure in transverse direction, as well as matrix or interface shear cracking, are taken into account. Computed failure probabilities are utilized in reducing cell stiffness based on the mesovolume concept. A numerical algorithm is developed predicting the damage evolution and deformation history of textile laminates. Effect of scatter of fiber orientation on cell properties is discussed. Weave influence on damage accumulation is illustrated with the help of an example of a Kevlar/epoxy laminate.

  9. Nonlinear effects on composite laminate thermal expansion

    NASA Technical Reports Server (NTRS)

    Hashin, Z.; Rosen, B. W.; Pipes, R. B.

    1979-01-01

    Analyses of Graphite/Polyimide laminates shown that the thermomechanical strains cannot be separated into mechanical strain and free thermal expansion strain. Elastic properties and thermal expansion coefficients of unidirectional Graphite/Polyimide specimens were measured as a function of temperature to provide inputs for the analysis. The + or - 45 degrees symmetric Graphite/Polyimide laminates were tested to obtain free thermal expansion coefficients and thermal expansion coefficients under various uniaxial loads. The experimental results demonstrated the effects predicted by the analysis, namely dependence of thermal expansion coefficients on load, and anisotropy of thermal expansion under load. The significance of time dependence on thermal expansion was demonstrated by comparison of measured laminate free expansion coefficients with and without 15 day delay at intermediate temperature.

  10. Fracture behavior of laminated discontinuously reinforced aluminum material

    SciTech Connect

    Osman, T.M. |; Lewandowski, J.J.; Lesuer, D.R.; Syn, C.K.; Hunt, W.H. Jr

    1994-05-01

    Laminated metallic composites are being developed for applications which require high specific stiffness and fracture resistance. Recent work with laminated discontinuously reinforced aluminum (DRA) materials has demonstrated the potential for marked improvements in stable crack growth resistance via extrinsic toughening. The purpose of this work is to compare the fracture mechanisms and fracture resistance of laminated DRA materials to unlaminated DRA materials. In particular, the production of extensive stable crack growth and the associated improvement in damage tolerance in DRA laminates is documented.

  11. Small Laminated Axial Turbine Design and Test Program.

    DTIC Science & Technology

    1980-12-01

    the Disk Rim During Startup and Shutdown 112 74 CME Rotor Burst Ratio 114 75 PCM Laminate Tool No. 21 115 76 Small Cruise Missile Laminated Turbine 117...Jr., H. R. Fisk and J. A. Vonada, ’Demonstration of a Cooled Laminated Integral Axial Turbine," AIAA Paper 77-949. Reprinted in Journal of Aircraft... Tooling (PC0) for the small diameter laminated rotor. 3 4. .< ,.C, DESIGN ANALYSIS Heat-Ttansfer Performance Predictions The expected metal temperature

  12. Ultrasonic transducer with laminated coupling wedge

    DOEpatents

    Karplus, Henry H. B.

    1976-08-03

    An ultrasonic transducer capable of use in a high-temperature environment incorporates a laminated metal coupling wedge including a reflecting edge shaped as a double sloping roof and a transducer crystal backed by a laminated metal sound absorber disposed so as to direct sound waves through the coupling wedge and into a work piece, reflections from the interface between the coupling wedge and the work piece passing to the reflecting edge. Preferably the angle of inclination of the two halves of the reflecting edge are different.

  13. Reliability analysis of continuous fiber composite laminates

    NASA Technical Reports Server (NTRS)

    Thomas, David J.; Wetherhold, Robert C.

    1991-01-01

    This paper describes two methods, the maximum distortion energy (MDE) and the principle of independent action (PIA), developed for the analysis of the reliability of a single continuous composite lamina. It is shown that, for the typical laminated plate structure, the individual lamina reliabilities can be combined in order to produce the upper and the lower bounds of reliability for the laminate, similar in nature to the bounds on properties produced from variational elastic methods. These limits were derived for both the interactive and the model failure considerations. Analytical expressions were also derived for the sensitivity of the reliability limits with respect to changes in the Weibull parameters and in loading conditions.

  14. Ultrahigh Carbon Steels and Their Laminates

    DTIC Science & Technology

    1990-02-01

    PROGRAM PROJECT TASK WORK UNIT 11LitME NT NO. No. NO. NO I I TITLE tiAtluda Seca.r.ty Ck~iaialasonJ Ultrahigh Carbon Steels and their laminates...PROM Aug. 1984 To- Fe~r--9O February 1, 1906 1S. SUPPLEMENTARY NOTATION Amore coinpetc tte of tie-program is: Low Density and Tough Steels with High...Hardenabihzty: Processing, Testing and Evaluation of UHC steels and their laminates 17 COSATI CODES Is.. SUBJECT TERMS (CoAtInai" on uvwrue iroleemary

  15. Glucotoxic conditions induce endoplasmic reticulum stress to cause caspase 3 mediated lamin B degradation in pancreatic β-cells: Protection by nifedipine

    PubMed Central

    Syeda, Khadija; Mohammed, Abiy M.; Arora, Daleep K.; Kowluru, Anjaneyulu

    2013-01-01

    Nuclear lamins form the lamina on the interior of the nuclear envelope, and are involved in the regulation of various cellular processes, including DNA replication and chromatin organization. Despite this evidence, little is known about potential alterations in nuclear metabolism, specifically lamin structure and integrity in isolated β-cells subjected to stress conditions, including chronic exposure to hyperglycemia [i.e., glucotoxicity]. Herein, we investigated effects of glucotoxic conditions on the catalytic activation of caspase 3 and the associated degradation of one of its substrate proteins, namely lamin-B. We report that incubation of insulin-secreting INS-1 832/13 cells, normal rat islets or human islets under glucotoxic conditions [20 mM; 12–48 hr] results in the degradation of native lamin B leading to accumulation of the degraded products in non-relevant cellular compartments, including cytosol. Moreover, the effects of high glucose on caspase 3 activation and lamin B degradation were mimicked by thapsigargin, a known inducer of endoplasmic reticulum stress [ER stress]. Nifedipine, a known blocker of calcium channel activation, inhibited high glucose-induced caspase 3 activation and lamin B degradation in these cells. 4-phenyl butyric acid, a known inhibitor of ER stress, markedly attenuated glucose-induced CHOP expression [ER stress marker], caspase 3 activation and lamin B degradation. We conclude that glucotoxic conditions promote caspase 3 activation and lamin B degradation, which may, in part, be due to increased ER stress under these conditions. We also provide further evidence to support beneficial effects of calcium channel blockers against metabolic dysfunction of the islet β-cell induced by hyperglycemic conditions. PMID:23994168

  16. Glucotoxic conditions induce endoplasmic reticulum stress to cause caspase 3 mediated lamin B degradation in pancreatic β-cells: protection by nifedipine.

    PubMed

    Syeda, Khadija; Mohammed, Abiy M; Arora, Daleep K; Kowluru, Anjaneyulu

    2013-11-01

    Nuclear lamins form the lamina on the interior of the nuclear envelope, and are involved in the regulation of various cellular processes, including DNA replication and chromatin organization. Despite this evidence, little is known about potential alterations in nuclear metabolism, specifically lamin structure and integrity in isolated β-cells subjected to stress conditions, including chronic exposure to hyperglycemia (i.e., glucotoxicity). Herein, we investigated effects of glucotoxic conditions on the catalytic activation of caspase 3 and the associated degradation of one of its substrate proteins, namely lamin-B. We report that incubation of insulin-secreting INS-1 832/13 cells, normal rat islets or human islets under glucotoxic conditions (20 mM; 12-48 h) results in the degradation of native lamin B leading to accumulation of the degraded products in non-relevant cellular compartments, including cytosol. Moreover, the effects of high glucose on caspase 3 activation and lamin B degradation were mimicked by thapsigargin, a known inducer of endoplasmic reticulum stress (ER stress). Nifedipine, a known blocker of calcium channel activation, inhibited high glucose-induced caspase 3 activation and lamin B degradation in these cells. 4-Phenyl butyric acid, a known inhibitor of ER stress, markedly attenuated glucose-induced CHOP expression (ER stress marker), caspase 3 activation and lamin B degradation. We conclude that glucotoxic conditions promote caspase 3 activation and lamin B degradation, which may, in part, be due to increased ER stress under these conditions. We also provide further evidence to support beneficial effects of calcium channel blockers against metabolic dysfunction of the islet β-cell induced by hyperglycemic conditions.

  17. Lamin A/C deficiency reduces circulating tumor cell resistance to fluid shear stress

    PubMed Central

    Denais, Celine; Chan, Maxine F.; Wang, Zhexiao; Lammerding, Jan

    2015-01-01

    Metastasis contributes to over 90% of cancer-related deaths and is initiated when cancer cells detach from the primary tumor, invade the basement membrane, and enter the circulation as circulating tumor cells (CTCs). While metastasis is viewed as an inefficient process with most CTCs dying within the bloodstream, it is evident that some CTCs are capable of resisting hemodynamic shear forces to form secondary tumors in distant tissues. We hypothesized that nuclear lamins A and C (A/C) act as key structural components within CTCs necessary to resist destruction from elevated shear forces of the bloodstream. Herein, we show that, compared with nonmalignant epithelial cells, tumor cells are resistant to elevated fluid shear forces in vitro that mimic those within the bloodstream, as evidenced by significant decreases in cellular apoptosis and necrosis. Knockdown of lamin A/C significantly reduced tumor cell resistance to fluid shear stress, with significantly increased cell death compared with parental tumor cell and nontargeting controls. Interestingly, lamin A/C knockdown increased shear stress-induced tumor cell apoptosis, but did not significantly affect cellular necrosis. These data demonstrate that lamin A/C is an important structural component that enables tumor cell resistance to fluid shear stress-mediated death in the bloodstream, and may thus facilitate survival and hematogenous metastasis of CTCs. PMID:26447202

  18. Evaluation of shear-compressive strength properties for laminated GFRP composites in electromagnet system

    NASA Astrophysics Data System (ADS)

    Song, Jun Hee; Kim, Hak Kun; Kim, Sam Yeon

    2014-07-01

    Laminated fiber-reinforced composites can be applied to an insulating structure of a nuclear fusion device. It is necessary to investigate the interlaminar fracture characteristics of the laminated composites for the assurance of design and structural integrity. The three methods used to prepare the glass fiber reinforced plastic composites tested in this study were vacuum pressure impregnation, high pressure laminate (HPL), and prepreg laminate. We discuss the design criteria for safe application of composites and the shear-compressive test methods for evaluating mechanical properties of the material. Shear-compressive tests could be performed successfully using series-type test jigs that were inclined 0°, 30°, 45°, 60°, and 75° to the normal axis. Shear strength depends strongly on the applied compressive stress. The design range of allowable shear stress was extended by use of the appropriate composite fabrication method. HPL had the largest design range, and the allowable interlaminar shear stress was 0.254 times the compressive stress.

  19. Multi-layer laminate structure and manufacturing method

    DOEpatents

    Keenihan, James R.; Cleereman, Robert J.; Eurich, Gerald; Graham, Andrew T.; Langmaid, Joe A.

    2013-01-29

    The present invention is premised upon a multi-layer laminate structure and method of manufacture, more particularly to a method of constructing the multi-layer laminate structure utilizing a laminate frame and at least one energy activated flowable polymer.

  20. Multi-layer laminate structure and manufacturing method

    DOEpatents

    Keenihan, James R [Midland, MI; Cleereman, Robert J [Midland, MI; Eurich, Gerald [Merrill, MI; Graham, Andrew T [Midland, MI; Langmaid, Joe A [Caro, MI

    2012-04-24

    The present invention is premised upon a multi-layer laminate structure and method of manufacture, more particularly to a method of constructing the multi-layer laminate structure utilizing a laminate frame and at least one energy activated flowable polymer.

  1. Impact performance of two bamboo-based laminated composites

    Treesearch

    Huanrong Liu; Zehui Jiang; Zhengjun Sun; Yan Yan; Zhiyong Cai; Xiubiao Zhang

    2017-01-01

    The present work aims to determine the impact performance of two bamboo-based laminated composites [bamboo/poplar laminated composite (BPLC) and bamboo/ glass fiber laminated composite (BGFLC)] using lowvelocity impact tests by a drop tower. In addition, fracture characteristics were evaluated using computed tomography (CT). Results showed that BPLC presented better...

  2. Simulating Progressive Damage of Notched Composite Laminates with Various Lamination Schemes

    NASA Astrophysics Data System (ADS)

    Mandal, B.; Chakrabarti, A.

    2017-05-01

    A three dimensional finite element based progressive damage model has been developed for the failure analysis of notched composite laminates. The material constitutive relations and the progressive damage algorithms are implemented into finite element code ABAQUS using user-defined subroutine UMAT. The existing failure criteria for the composite laminates are modified by including the failure criteria for fiber/matrix shear damage and delamination effects. The proposed numerical model is quite efficient and simple compared to other progressive damage models available in the literature. The efficiency of the present constitutive model and the computational scheme is verified by comparing the simulated results with the results available in the literature. A parametric study has been carried out to investigate the effect of change in lamination scheme on the failure behaviour of notched composite laminates.

  3. Flexural properties of glued-laminated Southern pine beams with laminations positioned by visual-stiffness criteria

    Treesearch

    R. C. Moody; Billy Bohannan

    1970-01-01

    To establish the effect of using modulus elasticity in addition to visual grade as criteria for the positioning of laminations in laminated beams, an experimental study on southern pine members was conducted. The beams were manufactured in accordance with current specifications for glued-laminated southern pine timber, except that (a) minimum-quality tension...

  4. Reciprocal knock-in mice to investigate the functional redundancy of lamin B1 and lamin B2

    PubMed Central

    Lee, John M.; Tu, Yiping; Tatar, Angelica; Wu, Daniel; Nobumori, Chika; Jung, Hea-Jin; Yoshinaga, Yuko; Coffinier, Catherine; de Jong, Pieter J.; Fong, Loren G.; Young, Stephen G.

    2014-01-01

    Lamins B1 and B2 (B-type lamins) have very similar sequences and are expressed ubiquitously. In addition, both Lmnb1- and Lmnb2-deficient mice die soon after birth with neuronal layering abnormalities in the cerebral cortex, a consequence of defective neuronal migration. The similarities in amino acid sequences, expression patterns, and knockout phenotypes raise the question of whether the two proteins have redundant functions. To investigate this topic, we generated “reciprocal knock-in mice”—mice that make lamin B2 from the Lmnb1 locus (Lmnb1B2/B2) and mice that make lamin B1 from the Lmnb2 locus (Lmnb2B1/B1). Lmnb1B2/B2 mice produced increased amounts of lamin B2 but no lamin B1; they died soon after birth with neuronal layering abnormalities in the cerebral cortex. However, the defects in Lmnb1B2/B2 mice were less severe than those in Lmnb1-knockout mice, indicating that increased amounts of lamin B2 partially ameliorate the abnormalities associated with lamin B1 deficiency. Similarly, increased amounts of lamin B1 in Lmnb2B1/B1 mice did not prevent the neurodevelopmental defects elicited by lamin B2 deficiency. We conclude that lamins B1 and B2 have unique roles in the developing brain and that increased production of one B-type lamin does not fully complement loss of the other. PMID:24672053

  5. Structures of the lamin A/C R335W and E347K mutants: Implications for dilated cardiolaminopathies

    SciTech Connect

    Bollati, Michela; Barbiroli, Alberto; Favalli, Valentina; Arbustini, Eloisa; Charron, Philippe; Bolognesi, Martino

    2012-02-10

    Highlights: Black-Right-Pointing-Pointer Defects in the Lamin AC gene have been linked to more than 10 laminopathies. Black-Right-Pointing-Pointer R335 and E347 are highly conserved residues, whose mutation preserves Coil2B 3D structure. Black-Right-Pointing-Pointer R335W and E347K mutations may affect LMNA interaction with proteins of nuclear lamina. -- Abstract: Dilated cardiomyopathy (DCM) is a condition whereby the normal muscular function of the myocardium is altered by specific or multiple aetiologies. About 25-35% of DCM patients show familial forms of the disease, with most mutations affecting genes encoding cytoskeletal proteins. Most of the DCM-related mutations fall in the Lamin AC gene, in particular in the Coil2B domain of the encoded protein. In this context, we focussed our studies on the crystal structures of two lamin Coil2B domain mutants (R335W and E347K). Both R335 and E347 are higly conserved residues whose substitution has little effects on the Coil2B domain three-dimensional structure; we can thus hypothesize that the mutations may interfere with the binding of components within the nuclear lamina, or of nuclear factors, that have been proposed to interact/associate with lamin A/C.

  6. Expression of nuclear membrane proteins in normal, hyperplastic, and neoplastic thyroid epithelial cells.

    PubMed

    Wang, Jieying; Kondo, Tetsuo; Yamane, Tetsu; Nakazawa, Tadao; Oish, Naoki; Mochizuki, Kunio; Katoh, Ryohei

    2015-10-01

    Emerin, lamin A/C, lamin B, and lamin-associated polypeptide 2 (LAP2) are nuclear membrane proteins that play an important role in maintaining nuclear structure and coordinating cell activity. We studied the expression and significance of nuclear membrane proteins in neoplastic thyroid cells by immunohistochemistry, RT-PCR, and real-time PCR. In papillary carcinomas (PCs), the nuclear proteins most frequently expressed at high levels were emerin (82 % positive), lamin A/C (64 %), and LAP2 (82 %). Follicular carcinomas (FCs) most frequently expressed lamin B, while none of the undifferentiated carcinomas (UCs) showed strong expression of emerin or lamin A/C. In all medullary carcinomas (MCs), intermediate to high levels of expression of lamin A/C and LAP2 were found. By RT-PCR analysis, messenger RNA (mRNA) expression of all nuclear membrane proteins except emerin was higher in PC than in normal tissue. Real-time PCR analysis showed that mRNA expression of nuclear membrane protein varied between cell lines. Our findings suggest that expression of nuclear membrane proteins may be related to follicular function in normal and hyperplastic follicles, and we hypothesize that they are also involved in the proliferation and differentiation of neoplastic thyroid cells. We suggest that they reflect the biological nature and/or function of normal, hyperplastic, and neoplastic thyroid cells and may have some value in diagnosing thyroid tumors.

  7. [Laminitis in cattle: a literature review].

    PubMed

    Lischer, C; Ossent, P

    1994-10-01

    Worldwide afflictions of the claws belong to the economically important diseases in dairy cattle. The significance of laminitis has gained importance in the last years since the condition is regarded as the most important predisposing factor for the development of lesions such as sole ulcer, white line disease and heel horn erosion. Apart from the clinical stages (acute, subacute, chronic, chronic-recurrent) there is also a subclinical form of laminitis which does not cause lameness. It is characterized by soft yellowish sole and heel horn with haemorrhages in the sole and along the white line. Laminitis is a multifactorial event in which nutrition, genetic disposition and the perinatal period, combined with the associated diseases of high-yielding cows, have a particular significance. Currently, two principally different hypotheses on the pathogenesis are discussed. The generally accepted theory bases on a disturbance in the microcirculation of the corium. According to the other theory the circulatory disturbances are secondary to changes which occur in the horn producing cells of the stratum basale of the epidermis. The predisposing factors and the pathogenesis of laminitis are discussed in the light of possible therapeutic and prophylactic measures.

  8. Laminated Root Rot of Western Conifers

    Treesearch

    E.E. Nelson; N.E. Martin; R.E. Williams

    1981-01-01

    Laminated root rot is caused by the native fungus Phellinus weirii (Murr.) Gilb. It occurs throughout the Northwestern United States and in southern British Columbia, Canada. The disease has also been reported in Japan and Manchuria. In the United States, the pathogen is most destructive in pure Douglas-fir stands west of the crest of the Cascade Range in Washington...

  9. Thermal postbuckling behavior of laminated composite plates

    SciTech Connect

    Singh, G.; Rao, G.V.; Lyengar, N.G.R. )

    1994-06-01

    Thermal buckling and postbuckling behavior of shear deformable laminated composite plates is investigated by employing a four-node rectangular C(sup 1) continuous finite element. The investigation reveals that the postbuckling path may not remain stable throughout. It is shown that secondary instabilities coupled with changes in the spatial deformation do take place from the postbuckling path. 15 refs.

  10. Thermal buckling of laminated composite plates

    SciTech Connect

    Chen, L.W.; Chen, L.Y.

    1987-01-01

    Thermal buckling of laminated composite plates subjected to a temperature change is studied. The displacement equations of equilibrium are used and Galerkin's method is employed to determine the critical buckling temperature. Clamped and simply supported boundary conditions are both considered. The effects of various parameters on the thermal buckling are examined. 18 references.

  11. Impermeable Robust Hydrogels via Hybrid Lamination.

    PubMed

    Parada, German A; Yuk, Hyunwoo; Liu, Xinyue; Hsieh, Alex J; Zhao, Xuanhe

    2017-07-17

    Hydrogels have been proposed for sensing, drug delivery, and soft robotics applications, yet most of these materials suffer from low mechanical robustness and high permeability to small molecules, limiting their widespread use. This study reports a general strategy and versatile method to fabricate robust, highly stretchable, and impermeable hydrogel laminates via hybrid lamination of an elastomer layer bonded between hydrogel layers. By controlling the layers' composition and thickness, it is possible to tune the stiffness of the impermeable hydrogels without sacrificing the stretchability. These hydrogel laminates exhibit ultralow surface coefficients of friction and, unlike common single-material hydrogels, do not allow diffusion of various molecules across the structure due to the presence of the elastomer layer. This feature is then used to release different model drugs and, in a subsequent experiment, to sense different pH conditions on the two sides of the hydrogel laminate. A potential healthcare application is shown using the presented method to coat medical devices (catheter, tubing, and condom) with hydrogel, to allow for drug release and sensing of environmental conditions for gastrointestinal or urinary tract. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Laminated insulators having heat dissipation means

    DOEpatents

    Niemann, R.C.; Mataya, K.F.; Gonczy, J.D.

    1980-04-24

    A laminated body is provided with heat dissipation capabilities. The insulator body is formed by dielectric layers interleaved with heat conductive layers, and bonded by an adhesive to form a composite structure. The heat conductive layers include provision for connection to an external thermal circuit.

  13. Laminated root rot in western North America.

    Treesearch

    Walter G. Thies; Rona N. Sturrock

    1995-01-01

    Laminated root rot, caused by Phellinus weirii (Murr.) Gilb., is a serious root disease affecting Douglas-fir and other commercially important species of conifers in northwestern North America. This report gives an overview of the dis-ease as it occurs in the Pacific Northwest in Canada and the United States. Information on recognizing crown...

  14. Doped LZO buffer layers for laminated conductors

    DOEpatents

    Paranthaman, Mariappan Parans [Knoxville, TN; Schoop, Urs [Westborough, MA; Goyal, Amit [Knoxville, TN; Thieme, Cornelis Leo Hans [Westborough, MA; Verebelyi, Darren T [Oxford, MA; Rupich, Martin W [Framingham, MA

    2010-03-23

    A laminated conductor includes a metallic substrate having a surface, a biaxially textured buffer layer supported by the surface of the substrate, the biaxially textured buffer layer comprising LZO and a dopant for mitigating metal diffusion through the LZO, and a biaxially textured conductor layer supported by the biaxially textured buffer layer.

  15. Micro-Cracking Detection in Laminated Composites

    NASA Technical Reports Server (NTRS)

    Chu, Tsuchin; Leyte, Alma; DiGregorio, Anthony; Russell, Samuel S.; Walker, James L.; Thom, Robert (Technical Monitor)

    2002-01-01

    Porosity and fatigue cracking are two critical factors that affect the performance and safety of cryogenic fuel tanks and feedlines made from unlined laminated or weaved carbon/epoxy materials. This paper presents the experiments to induce fatigue cracking of laminated composites through thermal cycling as well as the feasibility of using Thermography and Ultrasound Spectroscopy technology (UT) to detect and measure such micro-cracking. Carbon/epoxy laminated composite panels were built and cut into strips. These specimens were partially submerged in liquid nitrogen while subjected to various loads on a test machine. Edges of some specimens were polished and etched to determine the degree of micro-cracking. The rest of specimens were then examined with Thermography and Ultrasound Spectroscopy NDE systems to investigate the feasibility of finding such micro-cracking in the laminated composites. Thermography is utilized to determine changes in thermal diffusivity. The degree of cracking may reduce the apparent thermal diffusivity and therefore change the thermal response on the surface. Thermography testing was conducted on a group of specimens where it is desired to have some correlation between the predetermined stress and the thermography data. Ultrasound Spectroscopy was used to determine peak changes between the pre-stressed and stressed samples. Data from the inspections were analyzed and the results are presented in this paper.

  16. How to make a laminated diving board

    Treesearch

    U.S. Dept. of Agriculture. Forest Service. Forest Products Laboratory.

    1965-01-01

    The Forest Products Laboratory has developed a laminated diving board that has shown excellent performance characteristics. This board has given long, economical service under the severe moisture hazards and heavy service conditions such as found at public swimming places. The adhesive used is of the fully waterproof synthetic-resin type, which requires no protection...

  17. Fatigue damage development of various CFRP-laminates

    NASA Technical Reports Server (NTRS)

    Schulte, K.; Baron, CH.

    1988-01-01

    The chronic strength and fatigue behavior of a woven carbon-fiber reinforced laminate in a balanced eight-shaft satin weave style was compared to nonwoven laminates with an equivalent cross-ply layup. Half the fibers were arranged in the direction of the load and the other half perpendicular to it. Two types of nonwoven laminates consisting of continuous fibers and aligned discontinuous fibers, both produced from carbon fiber prepregs, were studied. The cross-ply laminate with continuous fiber showed the best characteristics with regard to both static strength and fatigue. The similarities and differences in damage mechanisms in the laminates are described.

  18. Nonlinear laminate analysis for metal matrix fiber composites

    NASA Technical Reports Server (NTRS)

    Chamis, C. C.; Sinclair, J. H.

    1981-01-01

    A nonlinear laminate analysis is described for predicting the mechanical behavior (stress-strain relationships) of angleplied laminates in which the matrix is strained nonlinearly by both the residual stress and the mechanical load and in which additional nonlinearities are induced due to progressive fiber fractures and ply relative rotations. The nonlinear laminate analysis (NLA) is based on linear composite mechanics and a piece wise linear laminate analysis to handle the nonlinear responses. Results obtained by using this nonlinear analysis on boron fiber/aluminum matrix angleplied laminates agree well with experimental data. The results shown illustrate the in situ ply stress-strain behavior and synergistic strength enhancement.

  19. Prediction of Composite Laminate Fracture: Micromechanics and Progressive Fracture

    NASA Technical Reports Server (NTRS)

    Gotsis, P. K.; Chamis, C. C.; Minnetyan, L.

    1996-01-01

    This report describes an investigation to predict first-ply failure and final fracture in selected composite laminates subjected to inplane loads. The laminates were composed of glass fiber and graphite fibers in epoxy matrices. Failure envelopes based on first-ply failure and laminate fracture were generated for combined loading of these laminates. Predictions were evaluated by micromechanics-based theory and progressive fracture. The results show that, for most cases, combined tensile loading significantly enhanced the laminate fracture stress in comparison to the uniaxial loading.

  20. Fatigue damage development of various CFRP-laminates

    NASA Technical Reports Server (NTRS)

    Schulte, K.; Baron, CH.

    1988-01-01

    The chronic strength and fatigue behavior of a woven carbon-fiber reinforced laminate in a balanced eight-shaft satin weave style was compared to nonwoven laminates with an equivalent cross-ply layup. Half the fibers were arranged in the direction of the load and the other half perpendicular to it. Two types of nonwoven laminates consisting of continuous fibers and aligned discontinuous fibers, both produced from carbon fiber prepregs, were studied. The cross-ply laminate with continuous fiber showed the best characteristics with regard to both static strength and fatigue. The similarities and differences in damage mechanisms in the laminates are described.

  1. Composite laminates with negative through-the-thickness Poisson's ratios

    NASA Technical Reports Server (NTRS)

    Herakovich, C. T.

    1984-01-01

    A simple analysis using two dimensional lamination theory combined with the appropriate three dimensional anisotropic constitutive equation is presented to show some rather surprising results for the range of values of the through-the-thickness effective Poisson's ratio nu sub xz for angle ply laminates. Results for graphite-epoxy show that the through-the-thickness effective Poisson's ratio can range from a high of 0.49 for a 90 laminate to a low of -0.21 for a + or - 25s laminate. It is shown that negative values of nu sub xz are also possible for other laminates.

  2. Composite laminates with negative through-the-thickness Poisson's ratios

    NASA Technical Reports Server (NTRS)

    Herakovich, C. T.

    1984-01-01

    A simple analysis using two-dimensional lamination theory combined with the appropriate three-dimensional anisotropic constitutive equation is presented to show some rather surprising results for the range of values of the through-the-thickness effective Poisson's ratio nu sub xz for angle ply laminates. Results for graphite-epoxy show that the through-the-thickness effective Poisson's ratio can range from a high of 0.49 for a 90 laminate to a low of -0.21 for a + or - 25s laminate. It is shown that negative values of nu sub xz are also possible for other laminates.

  3. Progressive delamination in polymer matrix composite laminates: A new approach

    NASA Technical Reports Server (NTRS)

    Chamis, C. C.; Murthy, P. L. N.; Minnetyan, L.

    1992-01-01

    A new approach independent of stress intensity factors and fracture toughness parameters has been developed and is described for the computational simulation of progressive delamination in polymer matrix composite laminates. The damage stages are quantified based on physics via composite mechanics while the degradation of the laminate behavior is quantified via the finite element method. The approach accounts for all types of composite behavior, laminate configuration, load conditions, and delamination processes starting from damage initiation, to unstable propagation, and to laminate fracture. Results of laminate fracture in composite beams, panels, plates, and shells are presented to demonstrate the effectiveness and versatility of this new approach.

  4. Compression failure of angle-ply laminates

    NASA Technical Reports Server (NTRS)

    Peel, Larry D.; Hyer, Michael W.; Shuart, Mark J.

    1991-01-01

    The present work deals with modes and mechanisms of failure in compression of angle-ply laminates. Experimental results were obtained from 42 angle-ply IM7/8551-7a specimens with a lay-up of ((plus or minus theta)/(plus or minus theta)) sub 6s where theta, the off-axis angle, ranged from 0 degrees to 90 degrees. The results showed four failure modes, these modes being a function of off-axis angle. Failure modes include fiber compression, inplane transverse tension, inplane shear, and inplane transverse compression. Excessive interlaminar shear strain was also considered as an important mode of failure. At low off-axis angles, experimentally observed values were considerably lower than published strengths. It was determined that laminate imperfections in the form of layer waviness could be a major factor in reducing compression strength. Previously developed linear buckling and geometrically nonlinear theories were used, with modifications and enhancements, to examine the influence of layer waviness on compression response. The wavy layer is described by a wave amplitude and a wave length. Linear elastic stress-strain response is assumed. The geometrically nonlinear theory, in conjunction with the maximum stress failure criterion, was used to predict compression failure and failure modes for the angle-ply laminates. A range of wave length and amplitudes were used. It was found that for 0 less than or equal to theta less than or equal to 15 degrees failure was most likely due to fiber compression. For 15 degrees less than theta less than or equal to 35 degrees, failure was most likely due to inplane transverse tension. For 35 degrees less than theta less than or equal to 70 degrees, failure was most likely due to inplane shear. For theta less than 70 degrees, failure was most likely due to inplane transverse compression. The fiber compression and transverse tension failure modes depended more heavily on wave length than on wave amplitude. Thus using a single

  5. On a generalized laminate theory with application to bending, vibration, and delamination buckling in composite laminates

    SciTech Connect

    Barbero, E.J.

    1989-01-01

    In this study, a computational model for accurate analysis of composite laminates and laminates with including delaminated interfaces is developed. An accurate prediction of stress distributions, including interlaminar stresses, is obtained by using the Generalized Laminate Plate Theory of Reddy in which layer-wise linear approximation of the displacements through the thickness is used. Analytical as well as finite-element solutions of the theory are developed for bending and vibrations of laminated composite plates for the linear theory. Geometrical nonlinearity, including buckling and postbuckling are included and used to perform stress analysis of laminated plates. A general two dimensional theory of laminated cylindrical shells is also developed in this study. Geometrical nonlinearity and transverse compressibility are included. Delaminations between layers of composite plates are modelled by jump discontinuity conditions at the interfaces. The theory includes multiple delaminations through the thickness. Geometric nonlinearity is included to capture layer buckling. The strain energy release rate distribution along the boundary of delaminations is computed by a novel algorithm. The computational models presented herein are accurate for global behavior and particularly appropriate for the study of local effects.

  6. Determining Shear Stress Distribution in a Laminate

    NASA Technical Reports Server (NTRS)

    Bednarcyk, Brett A.; Aboudi, Jacob; Yarrington, Phillip W.

    2010-01-01

    A "simplified shear solution" method approximates the through-thickness shear stress distribution within a composite laminate based on an extension of laminated beam theory. The method does not consider the solution of a particular boundary value problem; rather, it requires only knowledge of the global shear loading, geometry, and material properties of the laminate or panel. It is thus analogous to lamination theory in that ply-level stresses can be efficiently determined from global load resultants at a given location in a structure and used to evaluate the margin of safety on a ply-by-ply basis. The simplified shear solution stress distribution is zero at free surfaces, continuous at ply boundaries, and integrates to the applied shear load. The method has been incorporated within the HyperSizer commercial structural sizing software to improve its predictive capability for designing composite structures. The HyperSizer structural sizing software is used extensively by NASA to design composite structures. In the case of through-thickness shear loading on panels, HyperSizer previously included a basic, industry-standard, method for approximating the resulting shear stress distribution in sandwich panels. However, no such method was employed for solid laminate panels. The purpose of the innovation is to provide an approximation of the through-thickness shear stresses in a solid laminate given the through-thickness shear loads (Qx and Qy) on the panel. The method was needed for implementation within the HyperSizer structural sizing software so that the approximated ply-level shear stresses could be utilized in a failure theory to assess the adequacy of a panel design. The simplified shear solution method was developed based on extending and generalizing bi-material beam theory to plate-like structures. It is assumed that the through-thickness shear stresses arise due to local bending of the laminate induced by the through-thickness shear load, and by imposing

  7. Nuclear Physics in a biological context

    NASA Astrophysics Data System (ADS)

    Discher, Dennis

    2012-02-01

    A solid tissue can be soft like fat or brain, stiff like striated muscle and heart, or rigid like bone -- and of course every cell has a nucleus that contributes in some way small or large to tissue mechanics. Indeed, nuclei generally exhibit rheology and plasticity that reflects both the chromatin and the nuclear envelope proteins called lamins, all of which change in differentiation. Profiling of tissue nuclei shows that the nuclear intermediate filament protein Lamin-A/C varies over 30-fold between adult tissues and scales strongly with micro-elasticity of tissue, while other nuclear envelope components such as Lamin-B exhibit small variations. Lamin-A/C has been implicated in aging syndromes that affect muscle and fat but not brain, and we find nuclei in brain-derived cells are indeed dominated by Lamin-B and are much softer than nuclei derived from muscle cells with predominantly Lamin-A/C. In vitro, matrix elasticity can affect expression of nuclear envelope components in adult stem cells, and major changes in Lamin-A/C are indeed shown to direct lineage with lower levels favoring soft tissue and higher levels promoting rigid tissue lineage. Further molecular studies provide evidence that the nucleus transduces physical stress. References: (1) J.D. Pajerowski, K.N. Dahl, F.L. Zhong, P.J. Sammak, and D.E. Discher. Physical plasticity of the nucleus in stem cell differentiation. PNAS 104: 15619-15624 (2007). (2) A. Buxboim, I. Ivanova, and D.E. Discher. Matrix Elasticity, Cytoskeletal Forces, and Physics of the Nucleus: how deeply do cells `feel' outside and in? Journal of Cell Science 123: 297-308 (2010).

  8. Loss of lamin B1 results in prolongation of S phase and decondensation of chromosome territories

    PubMed Central

    Camps, Jordi; Wangsa, Darawalee; Falke, Martin; Brown, Markus; Case, Chanelle M.; Erdos, Michael R.; Ried, Thomas

    2014-01-01

    Nuclear lamin B1 (LMNB1) constitutes one of the major structural proteins in the lamina mesh. We silenced the expression of LMNB1 by RNA interference in the colon cancer cell line DLD-1 and showed a dramatic redistribution of H3K27me3 from the periphery to a more homogeneous nuclear dispersion. In addition, we observed telomere attrition and an increased frequency of micronuclei and nuclear blebs. By 3D-FISH analyses, we demonstrated that the volume and surface of chromosome territories were significantly larger in LMNB1-depleted cells, suggesting that LMNB1 is required to maintain chromatin condensation in interphase nuclei. These changes led to a prolonged S phase due to activation of Chk1. Finally, silencing of LMNB1 resulted in extensive changes in alternative splicing of multiple genes and in a higher number of enlarged nuclear speckles. Taken together, our results suggest a mechanistic role of the nuclear lamina in the organization of chromosome territories, maintenance of genome integrity and proper gene splicing.—Camps, J., Wangsa, D., Falke, M., Brown, M., Case, C. M., Erdos, M. R., Ried, T. Loss of lamin B1 results in prolongation of S phase and decondensation of chromosome territories. PMID:24732130

  9. Novel Remanufacturing Process of Recycled Polytetrafluoroethylene(PTFE)/GF Laminate

    NASA Astrophysics Data System (ADS)

    Xi, Z.; Ghita, O. R.; Johnston, P.; Evans, K. E.

    2011-01-01

    Currently, the PTFE/GF laminate and PTFE PCB manufacturers are under considerable pressure to address the recycling issues due to Waste Electrical and Electronic Equipment (WEEE) Directive, shortage of landfill capacity and cost of disposal. This study is proposing a novel manufacture method for reuse of the mechanical ground PTFE/Glass fibre (GF) laminate and production of the first reconstitute PTFE/GF laminate. The reconstitute PTFE/GF laminate proposed here consists of a layer of recycled sub-sheet, additional layers of PTFE and PTFE coated glass cloth, also covered by copper foils. The reconstitute PTFE/GF laminate showed good dielectric properties. Therefore, there is potential to use the mechanical ground PTFE/GF laminate powder to produce reconstitute PTFE/GF laminate, for use in high frequencies PCB applications.

  10. Tensile stress-strain behavior of boron/aluminum laminates

    NASA Technical Reports Server (NTRS)

    Sova, J. A.; Poe, C. C., Jr.

    1978-01-01

    The tensile stress-strain behavior of five types of boron/aluminum laminates was investigated. Longitudinal and transverse stress-strain curves were obtained for monotonic loading to failure and for three cycles of loading to successively higher load levels. The laminate strengths predicted by assuming that the zero deg plies failed first correlated well with the experimental results. The stress-strain curves for all the boron/aluminum laminates were nonlinear except at very small strains. Within the small linear regions, elastic constants calculated from laminate theory corresponded to those obtained experimentally to within 10 to 20 percent. A limited amount of cyclic loading did not affect the ultimate strength and strain for the boron/aluminum laminates. The laminates, however, exhibited a permanent strain on unloading. The Ramberg-Osgood equation was fitted to the stress-strain curves to obtain average curves for the various laminates.

  11. The innate immunity adaptor SARM translocates to the nucleus to stabilize lamins and prevent DNA fragmentation in response to pro-apoptotic signaling.

    PubMed

    Sethman, Chad R; Hawiger, Jacek

    2013-01-01

    Sterile alpha and armadillo-motif containing protein (SARM), a highly conserved and structurally unique member of the MyD88 family of Toll-like receptor adaptors, plays an important role in innate immunity signaling and apoptosis. Its exact mechanism of intracellular action remains unclear. Apoptosis is an ancient and ubiquitous process of programmed cell death that results in disruption of the nuclear lamina and, ultimately, dismantling of the nucleus. In addition to supporting the nuclear membrane, lamins serve important roles in chromatin organization, epigenetic regulation, transcription, nuclear transport, and mitosis. Mutations and other damage that destabilize nuclear lamins (laminopathies) underlie a number of intractable human diseases. Here, we report that SARM translocates to the nucleus of human embryonic kidney cells by using its amino-terminal Armadillo repeat region. Within the nucleus, SARM forms a previously unreported lattice akin to the nuclear lamina scaffold. Moreover, we show that SARM protects lamins from apoptotic degradation and reduces internucleosomal DNA fragmentation in response to signaling induced by the proinflammatory cytokine Tumor Necrosis Factor alpha. These findings indicate an important link between the innate immunity adaptor SARM and stabilization of nuclear lamins during inflammation-driven apoptosis in human cells.

  12. Novel origin of lamin-derived cytoplasmic intermediate filaments in tardigrades

    PubMed Central

    Hering, Lars; Bouameur, Jamal-Eddine; Reichelt, Julian; Magin, Thomas M; Mayer, Georg

    2016-01-01

    Intermediate filament (IF) proteins, including nuclear lamins and cytoplasmic IF proteins, are essential cytoskeletal components of bilaterian cells. Despite their important role in protecting tissues against mechanical force, no cytoplasmic IF proteins have been convincingly identified in arthropods. Here we show that the ancestral cytoplasmic IF protein gene was lost in the entire panarthropod (onychophoran + tardigrade + arthropod) rather than arthropod lineage and that nuclear, lamin-derived proteins instead acquired new cytoplasmic roles at least three times independently in collembolans, copepods, and tardigrades. Transcriptomic and genomic data revealed three IF protein genes in the tardigrade Hypsibius dujardini, one of which (cytotardin) occurs exclusively in the cytoplasm of epidermal and foregut epithelia, where it forms belt-like filaments around each epithelial cell. These results suggest that a lamin derivative has been co-opted to enhance tissue stability in tardigrades, a function otherwise served by cytoplasmic IF proteins in all other bilaterians. DOI: http://dx.doi.org/10.7554/eLife.11117.001 PMID:26840051

  13. Expansion of the Nucleoplasmic Reticulum Requires the Coordinated Activity of Lamins and CTP:Phosphocholine Cytidylyltransferase α

    PubMed Central

    Gehrig, Karsten; Cornell, Rosemary B.

    2008-01-01

    The nucleoplasmic reticulum (NR), a nuclear membrane network implicated in signaling and transport, is formed by the biosynthetic and membrane curvature-inducing properties of the rate-limiting enzyme in phosphatidylcholine synthesis, CTP:phosphocholine cytidylyltransferase (CCT) α. The NR is formed by invagination of the nuclear envelope and has an underlying lamina that may contribute to membrane tubule formation or stability. In this study we investigated the role of lamins A and B in NR formation in response to expression and activation of endogenous and fluorescent protein-tagged CCTα. Similarly to endogenous CCTα, CCT-green fluorescent protein (GFP) reversibly translocated to nuclear tubules projecting from the NE in response to oleate, a lipid promoter of CCT membrane binding. Coexpression and RNA interference experiments revealed that both CCTα and lamin A and B were necessary for NR proliferation. Expression of CCT-GFP mutants with compromised membrane-binding affinity produced fewer nuclear tubules, indicating that the membrane-binding function of CCTα promotes the expansion of the NR. Proliferation of atypical bundles of nuclear membrane tubules by a CCTα mutant that constitutively associated with membranes revealed that expansion of the double-bilayer NR requires the coordinated assembly of an underlying lamin scaffold and induction of membrane curvature by CCTα. PMID:17959832

  14. Multi-Scale Creep Analysis of Plain-Woven Laminates Using Time-Dependent Homogenization Theory:. Effects of Laminate Configuration

    NASA Astrophysics Data System (ADS)

    Nakata, K.; Matsuda, T.; Kawai, M.

    In this study, multi-scale creep analysis of plain-woven GFRP laminates is performed using the time-dependent homogenization theory developed by the present authors. First, point-symmetry of internal structures of plain-woven laminates is utilized for a boundary condition of unit cell problems, reducing the domain of analysis to 1/4 and 1/8 for in-phase and out-of-phase laminate configurations, respectively. The time-dependent homogenization theory is then reconstructed for these domains of analysis. Using the present method, in-plane creep behavior of plain-woven glass fiber/epoxy laminates subjected to a constant stress is analyzed. The results are summarized as follows: (1) The in-plane creep behavior of the plain-woven GFRP laminates exhibits marked anisotropy. (2) The laminate configurations considerably affect the creep behavior of the laminates.

  15. Hutchinson–Gilford progeria mutant lamin A primarily targets human vascular cells as detected by an anti-Lamin A G608G antibody

    PubMed Central

    McClintock, Dayle; Gordon, Leslie B.; Djabali, Karima

    2006-01-01

    Hutchinson–Gilford progeria syndrome (HGPS; Online Mendelian Inheritance in Man accession no. 176670) is a rare disorder that is characterized by segmental premature aging and death between 7 and 20 years of age from severe premature atherosclerosis. Mutations in the LMNA gene are responsible for this syndrome. Approximately 80% of HGPS cases are caused by a G608 (GGC→GGT) mutation within exon 11 of LMNA, which elicits a deletion of 50 aa near the C terminus of prelamin A. In this article, we present evidence that the mutant lamin A (progerin) accumulates in the nucleus in a cellular age-dependent manner. In human HGPS fibroblast cultures, we observed, concomitantly to nuclear progerin accumulation, severe nuclear envelope deformations and invaginations preventable by farnesyltransferase inhibition. Nuclear alterations affect cell-cycle progression and cell migration and elicit premature senescence. Strikingly, skin biopsy sections from a subject with HGPS showed that the truncated lamin A accumulates primarily in the nuclei of vascular cells. This finding suggests that accumulation of progerin is directly involved in vascular disease in progeria. PMID:16461887

  16. Hutchinson-Gilford progeria mutant lamin A primarily targets human vascular cells as detected by an anti-Lamin A G608G antibody.

    PubMed

    McClintock, Dayle; Gordon, Leslie B; Djabali, Karima

    2006-02-14

    Hutchinson-Gilford progeria syndrome (HGPS; Online Mendelian Inheritance in Man accession no. 176670) is a rare disorder that is characterized by segmental premature aging and death between 7 and 20 years of age from severe premature atherosclerosis. Mutations in the LMNA gene are responsible for this syndrome. Approximately 80% of HGPS cases are caused by a G608 (GGC-->GGT) mutation within exon 11 of LMNA, which elicits a deletion of 50 aa near the C terminus of prelamin A. In this article, we present evidence that the mutant lamin A (progerin) accumulates in the nucleus in a cellular age-dependent manner. In human HGPS fibroblast cultures, we observed, concomitantly to nuclear progerin accumulation, severe nuclear envelope deformations and invaginations preventable by farnesyltransferase inhibition. Nuclear alterations affect cell-cycle progression and cell migration and elicit premature senescence. Strikingly, skin biopsy sections from a subject with HGPS showed that the truncated lamin A accumulates primarily in the nuclei of vascular cells. This finding suggests that accumulation of progerin is directly involved in vascular disease in progeria.

  17. Caspase-3 is required in the apoptotic disintegration of the nuclear matrix

    SciTech Connect

    Kivinen, Katri; Kallajoki, Markku; Taimen, Pekka . E-mail: pekka.taimen@utu.fi

    2005-11-15

    Apoptotic breakdown of cellular structures is largely mediated by caspases. One target of degradation is a proteinaceous framework of the nucleus termed the nuclear matrix. We compared the apoptotic changes of the nuclear matrix in staurosporine-treated caspase-3-deficient MCF-7 cells transfected with intact CASP-3 gene (MCF-7c3) or an empty vector (MCF-7v) as a control. Nuclear Mitotic Apparatus protein (NuMA), lamin A/C and lamin B were used as markers for internal nuclear matrix and peripheral nuclear lamina, respectively. In both cell lines, staurosporine induced rapid cytoplasmic shrinkage and partial chromatin condensation. MCF-7c3 cells formed apoptotic bodies, whereas MCF-7v cells did not. NuMA and lamins were actively cleaved in MCF-7c3 cells following caspase-3 activation, but only minimal or no cleavage was detected in MCF-7v cells. Interestingly, lamin B but not lamin A/C was relocated into cytoplasmic granules in apoptotic MCF-7v cells. Pancaspase inhibitor, z-VAD-fmk, prevented the apoptotic changes, while caspase-3 inhibitor, z-DEVD-fmk, induced lamin B granules in both cell lines. These results show that caspase-3 is involved in the cleavage of NuMA and lamins either directly or by activating other proteases. This may be essential for disintegration of the nuclear structure during apoptosis.

  18. Technical specifications for the successful fabrication of laminated seismic isolation bearings

    SciTech Connect

    Kulak, R.F.

    1992-05-01

    High damping laminated elastomeric bearings are becoming one of the preferred devices for isolating large buildings and structures. IN the United States, the current reference design for the Advanced Liquid Metal Reactor uses laminated bearings for seismic isolation. These bearing are constructed from alternating layers of rubber and steel plates. They are typically designed for shear strains between 50 to 100 percent and expected to sustain two to three times these levels for beyond design basis loading considerations. The technical specifications used to procure these bearings are an important factor in assuring that the bearings that are installed under nuclear structures meet the performance requirements of the design. The key aspects of the current version of the Technical Specifications are discussed in this paper.

  19. Technical specifications for the successful fabrication of laminated seismic isolation bearings

    SciTech Connect

    Kulak, R.F.

    1992-01-01

    High damping laminated elastomeric bearings are becoming one of the preferred devices for isolating large buildings and structures. IN the United States, the current reference design for the Advanced Liquid Metal Reactor uses laminated bearings for seismic isolation. These bearing are constructed from alternating layers of rubber and steel plates. They are typically designed for shear strains between 50 to 100 percent and expected to sustain two to three times these levels for beyond design basis loading considerations. The technical specifications used to procure these bearings are an important factor in assuring that the bearings that are installed under nuclear structures meet the performance requirements of the design. The key aspects of the current version of the Technical Specifications are discussed in this paper.

  20. Micromechanical Modeling of Impact Damage Mechanisms in Unidirectional Composite Laminates

    NASA Astrophysics Data System (ADS)

    Meng, Qinghua; Wang, Zhenqing

    2016-12-01

    Composite laminates are susceptible to the transverse impact loads resulting in significant damage such as matrix cracking, fiber breakage and delamination. In this paper, a micromechanical model is developed to predict the impact damage of composite laminates based on microstructure and various failure models of laminates. The fiber and matrix are represented by the isotropic and elastic-plastic solid, and their impact failure behaviors are modeled based on shear damage model. The delaminaton failure is modeling by the interface element controlled by cohesive damage model. Impact damage mechanisms of laminate are analyzed by using the micromechanical model proposed. In addition, the effects of impact energy and laminated type on impact damage behavior of laminates are investigated. Due to the damage of the surrounding matrix near the impact point caused by the fiber deformation, the surface damage area of laminate is larger than the area of ​​impact projectile. The shape of the damage area is roughly rectangle or elliptical with the major axis extending parallel to the fiber direction in the surface layer of laminate. The alternating laminated type with two fiber directions is more propitious to improve the impact resistance of laminates.

  1. The structural response of unsymmetrically laminated composite cylinders

    NASA Technical Reports Server (NTRS)

    Butler, T. A.; Hyer, M. W.

    1989-01-01

    The responses of an unsymmetrically laminated fiber-reinforced composite cylinder to an axial compressive load, a torsional load, and the temperature change associated with cooling from the processing temperature to the service temperature are investigated. These problems are considered axisymmetric and the response is studied in the context of linear elastic material behavior and geometrically linear kinematics. Four different laminates are studied: a general unsymmetric laminate; two unsymmetric but more conventional laminates; and a conventional quasi-isotropic symmetric laminate. The responses based on closed-form solutions for different boundary conditions are computed and studied in detail. Particular emphasis is directed at understanding the influence of elastic couplings in the laminates. The influence of coupling decreased from a large effect in the general unsymmetric laminate, to practically no effect in the quasi-isotropic laminate. For example, the torsional loading of the general unsymmetric laminate resulted in a radial displacement. The temperature change also caused a significant radial displacement to occur near the ends of the cylinder. On the other hand, the more conventional unsymmetric laminate and the quasi-isotropic cylinder did not deform radially when subjected to a torsional load. From the results obtained, it is clear the degree of elastic coupling can be controlled and indeed designed into a cylinder, the degree and character of the coupling being dictated by the application.

  2. Thermomechanical postbuckling analysis of laminated composite shells

    SciTech Connect

    Averill, R.C.; Reddy, J.N.

    1993-01-01

    The nonlinear response of laminated composite structures subjected to thermal loads is investigated. Analysis is performed using a refined theory and an associated finite element model for geometrically nonlinear analysis of laminated composite shell structures. The model is based on a third-order displacement field which accounts for both transverse shear and transverse normal deformations. Numerical studies of simply-supported plates and cylindrical panels indicate that when the panels are free to expand or contract in the transverse direction, the predicted critical buckling temperatures do not depend significantly upon whether or not transverse normal deformations are explicitly accounted for in the analysis model. However, the critical buckling temperatures are strongly dependent upon whether or not the transverse normal deformations are restrained along the boundaries of the panels. 25 refs.

  3. Fabrication of CFRP/Al Active Laminates

    NASA Astrophysics Data System (ADS)

    Asanuma, Hiroshi; Haga, Osamu; Ohira, Junichiro; Takemoto, Kyosuke; Imori, Masataka

    This paper describes fabrication and evaluation of the active laminate. It was made by hot-pressing of an aluminum plate as a high CTE material, a unidirectional CFRP prepreg as a low CTE material and an electric resistance heater, a KFRP prepreg as a low CTE material and an insulator between them, and copper foils as electrodes. In this study, fabricating conditions and performances such as curvature change and output force were examined. Under optimized fabricating conditions, it became clear that 1) the curvature of the active laminate linearly changes as a function of temperature, between room temperature and its hot pressing temperature without hysteresis by electric resistance heating of carbon fiber in the CFRP layer and cooling, and 2) the output force against a fixed punch almost linearly increases with increasing temperature during heating from 313K up to around the glass transition temperature of the epoxy matrix.

  4. Thermal stresses in thick laminated composite shells

    NASA Technical Reports Server (NTRS)

    Yuan, F. G.

    1993-01-01

    The paper provides an analytical formulation to investigate the thermomechanical behavior of thick composite shells subjected to a temperature distribution which varies arbitrarily in the radial direction. For illustrative purposes, shells under uniform temperature change are presented. It is found that thermal twist would occur even for symmetric laminated shells. Under uniform temperature rise, results for off-axis graphite/epoxy shells show that extensional-shear coupling can cause tensile radial stress throughout the shell and tensile hoop stress in the inner region. Laminated graphite/epoxy shells can exhibit negative effective thermal expansion coefficients in the longitudinal and transverse directions. Finally, the stacking sequence has a strong influence on the thermal stress distributions.

  5. High energy impact on woven laminates

    NASA Astrophysics Data System (ADS)

    López-Puente, J.; Zaera, R.; Navarro, C.

    2003-09-01

    The influence of high velocity impacts on CFRPs was studied by launching Spherical steel masses, at velocities from 60 m/s to 550 m/s, against carbon fiber/epoxy woven laminates. The extension of the damage induced in the laminate was measured by C-Scan. Finite element numerical simulation of the impact test used a failure model based on the Chang-Chang model. A comparison was made of the damaged areas resulting from non-destructive inspection of the specimens and those predicted by numerical simulation. To conclue the analysis, an analytical model developed by Cantwell-Morton was used to calculate the residual velocity of the projectile after perforation. The residual velocities predicted by numerical and by analytical models, were also compared.

  6. Investigating Delamination Migration in Composite Tape Laminates

    NASA Technical Reports Server (NTRS)

    Ratcliffe, James G.; DeCarvalho, Nelson V.

    2014-01-01

    A modification to a recently developed test specimen designed to investigate migration of a delamination between neighboring ply interfaces in tape laminates is presented. The specimen is a cross-ply laminated beam consisting of 40 plies with a polytetrafluoroethylene insert spanning part way along its length. The insert is located between a lower 0-degree ply (specimen length direction) and a stack of four 90-degree plies (specimen width direction). The modification involved a stacking sequence that promotes stable delamination growth prior to migration, and included a relocation of the insert from the specimen midplane to the interface between plies 14 and 15. Specimens were clamped at both ends onto a rigid baseplate and loaded on their upper surface via a piano hinge assembly, resulting in a predominantly flexural loading condition. Tests were conducted with the load-application point positioned at various locations along a specimen's span. This position affected the sequence of damage events during a test.

  7. Fatigue Damage Mechanisms in Advanced Hybrid Titanium Composite Laminates

    NASA Technical Reports Server (NTRS)

    Johnson, W. Steven; Rhymer, Donald W.; St.Clair, Terry L. (Technical Monitor)

    2000-01-01

    Hybrid Titanium Composite Laminates (HTCL) are a type of hybrid composite laminate with promise for high-speed aerospace applications, specifically designed for improved damage tolerance and strength at high-temperature (350 F, 177 C). However, in previous testing, HTCL demonstrated a propensity to excessive delamination at the titanium/PMC interface following titanium cracking. An advanced HTCL has been constructed with an emphasis on strengthening this interface, combining a PETI-5/IM7 PMC with Ti-15-3 foils prepared with an alkaline-perborate surface treatment. This paper discusses how the fatigue capabilities of the "advanced" HTCL compare to the first generation HTCL which was not modified for interface optimization, in both tension-tension (R = 0.1) and tension-compression (R=-0.2). The advanced HTCL under did not demonstrate a significant improvement in fatigue life, in either tension-tension or tension-compression loading. However, the advanced HTCL proved much more damage tolerant. The R = 0.1 tests revealed the advanced HTCL to increase the fatigue life following initial titanium ply damage up to 10X that of the initial HTCL at certain stress levels. The damage progression following the initial ply damage demonstrated the effect of the strengthened PMC/titanium interface. Acetate film replication of the advanced HTCL edges showed a propensity for some fibers in the adjacent PMC layers to fail at the point of titanium crack formation, suppressing delamination at the Ti/PMC interface. The inspection of failure surfaces validated these findings, revealing PMC fibers bonded to the majority of the titanium surfaces. Tension compression fatigue (R = -0.2) demonstrated the same trends in cycles between initial damage and failure, damage progression, and failure surfaces. Moreover, in possessing a higher resistance to delamination, the advanced HTCL did not exhibit buckling following initial titanium ply cracking under compression unlike the initial HTCL.

  8. The B-type lamin is required for somatic repression of testis-specific gene clusters

    PubMed Central

    Shevelyov, Y. Y.; Lavrov, S. A.; Mikhaylova, L. M.; Nurminsky, I. D.; Kulathinal, R. J.; Egorova, K. S.; Rozovsky, Y. M.; Nurminsky, D. I.

    2009-01-01

    Large clusters of coexpressed tissue-specific genes are abundant on chromosomes of diverse species. The genes coordinately misexpressed in diverse diseases are also found in similar clusters, suggesting that evolutionarily conserved mechanisms regulate expression of large multigenic regions both in normal development and in its pathological disruptions. Studies on individual loci suggest that silent clusters of coregulated genes are embedded in repressed chromatin domains, often localized to the nuclear periphery. To test this model at the genome-wide scale, we studied transcriptional regulation of large testis-specific gene clusters in somatic tissues of Drosophila. These gene clusters showed a drastic paucity of known expressed transgene insertions, indicating that they indeed are embedded in repressed chromatin. Bioinformatics analysis suggested the major role for the B-type lamin, LamDmo, in repression of large testis-specific gene clusters, showing that in somatic cells as many as three-quarters of these clusters interact with LamDmo. Ablation of LamDmo by using mutants and RNAi led to detachment of testis-specific clusters from nuclear envelope and to their selective transcriptional up-regulation in somatic cells, thus providing the first direct evidence for involvement of the B-type lamin in tissue-specific gene repression. Finally, we found that transcriptional activation of the lamina-bound testis-specific gene cluster in male germ line is coupled with its translocation away from the nuclear envelope. Our studies, which directly link nuclear architecture with coordinated regulation of tissue-specific genes, advance understanding of the mechanisms underlying both normal cell differentiation and developmental disorders caused by lesions in the B-type lamins and interacting proteins. PMID:19218438

  9. Digital holographic nondestructive testing of laminate composite

    NASA Astrophysics Data System (ADS)

    Karray, Mayssa; Christophe, Poilane; Gargouri, Mohamed; Picart, Pascal

    2016-09-01

    Optical digital holographic techniques can be used for nondestructive testing of materials. Digital holographic nondestructive testing essentially measures deformations on the surface of the object. However, there is sufficient sensitivity to detect subsurface and internal defects in metallic and composite specimens. We investigate and discuss the vibration analysis of laminated composite glass-epoxy using time averaging in digital Fresnel holography to visualize the modes of vibration and to test the integrity of the structures of studied materials.

  10. Laminated grid and web magnetic cores

    DOEpatents

    Sefko, John; Pavlik, Norman M.

    1984-01-01

    A laminated magnetic core characterized by an electromagnetic core having core legs which comprise elongated apertures and edge notches disposed transversely to the longitudinal axis of the legs, such as high reluctance cores with linear magnetization characteristics for high voltage shunt reactors. In one embodiment the apertures include compact bodies of microlaminations for more flexibility and control in adjusting permeability and/or core reluctance.

  11. Examination of Blistering of Fiberglass Reinforced Laminates

    DTIC Science & Technology

    1987-06-26

    laminates produced by non-standard methods would give a different ’signature’ than those made normally. I AGA Vision 688 Inframetrics 210 Spectral Range 2 to...Resolution .2C .40C and .20C Test Observations The observed output from the AGA unit appeared to be superior to I that of the Inframetrics . The resolution...definite using the Inframetrics unit. The fiberglass reinforced plastic panels (FRP) were positioned f vertically, approximately 15 feet from the infrared

  12. Extended Flow Life Laminating Resin System

    DTIC Science & Technology

    1976-06-01

    respo ^cerntrt ^^«üi-C^Äl. "E». How Life Laminating Resin System". The work was performed under the ^f^^L^l^L^tterl^rHr^ Laboratory (AFWL/ HBC ) Air...450 psi) CF) (from TMA) N/A N/A 340 240 230 320 310 470 455 385 390 365 400 300 310 (390)*** 375 (430)*** *0estructi ve exotherm

  13. Nondimensional impact models for composite laminates

    NASA Technical Reports Server (NTRS)

    Sankar, B. V.; Nguyen, P. T.; Ku, C.

    1990-01-01

    The equations governing the problem of low-velocity impact of a simply supported rectangular laminated plate are nondimensionalized such that the problem is defined in terms of five dimensionless parameters. A parametric study using the Graeco-Latin Factorial Plan is performed. Semi-empirical formulas for maximum impact force, impact duration, and maximum back surface strains are obtained. It is found that some of the simple impact models provide the bounds for the case of impact on a finite extent plate.

  14. Residual stresses in polymer matrix composite laminates

    NASA Technical Reports Server (NTRS)

    Hahn, H. T.

    1976-01-01

    Residual stresses in composites are induced during fabrication and by environmental exposure. The theory formulated can describe the shrinkage commonly observed after a thermal expansion test. Comparison between the analysis and experimental data for laminates of various material systems indicates that the residual stress-free temperature can be lower than the curing temperature, depending on the curing process. Effects of residual stresses on ply failure including the acoustic emission characteristics are discussed.

  15. Delamination stresses in semicircular laminated composite bars

    NASA Technical Reports Server (NTRS)

    Ko, William L.

    1988-01-01

    Using anisotropic elasticity theory, delamination stresses in a semicircular laminated composite curved bar subjected to end forces and end moments were calculated, and their radial locations determined. A family of design curves was presented, showing variation of the intensity of delamination stresses and their radial locations with different geometry and different degrees of anisotropy of the curved bar. The effect of anisotropy on the location of peak delamination stress was found to be small.

  16. The Novel Nuclear Envelope Protein KAKU4 Modulates Nuclear Morphology in Arabidopsis[W

    PubMed Central

    Goto, Chieko; Tamura, Kentaro; Fukao, Yoichiro; Shimada, Tomoo; Hara-Nishimura, Ikuko

    2014-01-01

    In animals, the nuclear lamina is a fibrillar meshwork on the inner surface of the nuclear envelope, composed of coiled-coil lamin proteins and lamin binding membrane proteins. Plants also have a meshwork on the inner surface of the nuclear envelope, but little is known about its composition other than the presence of members of the CROWDED NUCLEI (CRWN) protein family, possible plant lamin analogs. Here, we describe a candidate lamina component, based on two Arabidopsis thaliana mutants (kaku2 and kaku4) with aberrant nuclear morphology. The responsible gene in kaku2 encodes CRWN1, and the responsible gene in kaku4 encodes a plant-specific protein of unknown function (KAKU4) that physically interacts with CRWN1 and its homolog CRWN4. Immunogold labeling revealed that KAKU4 localizes at the inner nuclear membrane. KAKU4 deforms the nuclear envelope in a dose-dependent manner, in association with nuclear membrane invagination and stack formation. The KAKU4-dependent nuclear envelope deformation was enhanced by overaccumulation of CRWN1, although KAKU4 can deform the nuclear envelope even in the absence of CRWN1 and/or CRWN4. Together, these results suggest that plants have evolved a unique lamina-like structure to modulate nuclear shape and size. PMID:24824484

  17. Modeling Composite Laminate Crushing for Crash Analysis

    NASA Technical Reports Server (NTRS)

    Fleming, David C.; Jones, Lisa (Technical Monitor)

    2002-01-01

    Crash modeling of composite structures remains limited in application and has not been effectively demonstrated as a predictive tool. While the global response of composite structures may be well modeled, when composite structures act as energy-absorbing members through direct laminate crushing the modeling accuracy is greatly reduced. The most efficient composite energy absorbing structures, in terms of energy absorbed per unit mass, are those that absorb energy through a complex progressive crushing response in which fiber and matrix fractures on a small scale dominate the behavior. Such failure modes simultaneously include delamination of plies, failure of the matrix to produce fiber bundles, and subsequent failure of fiber bundles either in bending or in shear. In addition, the response may include the significant action of friction, both internally (between delaminated plies or fiber bundles) or externally (between the laminate and the crushing surface). A figure shows the crushing damage observed in a fiberglass composite tube specimen, illustrating the complexity of the response. To achieve a finite element model of such complex behavior is an extremely challenging problem. A practical crushing model based on detailed modeling of the physical mechanisms of crushing behavior is not expected in the foreseeable future. The present research describes attempts to model composite crushing behavior using a novel hybrid modeling procedure. Experimental testing is done is support of the modeling efforts, and a test specimen is developed to provide data for validating laminate crushing models.

  18. Laminate analogy for composites enhanced concrete structures

    SciTech Connect

    Chamis, C.C.; Gotsis, P.K.

    1997-10-01

    A new and effective method is described to design composites to repair damage or enhance the overload strength of concrete infrastructures. The method is based on laminate analogy which is derivable from composite mechanics and available in computer codes. It is used to simulate structural sections made from reinforced concrete which are typical in infrastructures as well as select reinforced concrete structures. The structural sections are represented by a number of layers through the thickness where different layers are used for the concrete, for the reinforcing steel in concrete, and for the composite. The reinforced concrete structures are represented with finite elements where the element stiffness parameters are from the structural sections which are represented by the laminate analogy. The load carrying capability of the structure is determined by progressive structural fracture. Results show up to 40 percent improvements for damage and for overload enhancement with relatively small laminate thickness for the structural sections and up to three times for the composite enhanced select structures (arch and dome).

  19. Superconductivity in Ca-doped graphene laminates

    PubMed Central

    Chapman, J.; Su, Y.; Howard, C. A.; Kundys, D.; Grigorenko, A. N.; Guinea, F.; Geim, A. K.; Grigorieva, I. V.; Nair, R. R.

    2016-01-01

    Despite graphene’s long list of exceptional electronic properties and many theoretical predictions regarding the possibility of superconductivity in graphene, its direct and unambiguous experimental observation has not been achieved. We searched for superconductivity in weakly interacting, metal decorated graphene crystals assembled into so-called graphene laminates, consisting of well separated and electronically decoupled graphene crystallites. We report robust superconductivity in all Ca-doped graphene laminates. They become superconducting at temperatures (Tc) between ≈4 and ≈6 K, with Tc’s strongly dependent on the confinement of the Ca layer and the induced charge carrier concentration in graphene. We find that Ca is the only dopant that induces superconductivity in graphene laminates above 1.8 K among several dopants used in our experiments, such as potassium, caesium and lithium. By revealing the tunability of the superconducting response through doping and confinement of the metal layer, our work shows that achieving superconductivity in free-standing, metal decorated monolayer graphene is conditional on an optimum confinement of the metal layer and sufficient doping, thereby bringing its experimental realization within grasp. PMID:26979564

  20. Laminate articles on biaxially textured metal substrates

    DOEpatents

    Beach, David B.; Morrell, Jonathan S.; Paranthaman, Mariappan; Chirayil, Thomas; Specht, Eliot D.; Goyal, Amit

    2003-12-16

    A laminate article comprises a substrate and a biaxially textured (RE.sup.1.sub.x RE.sup.2.sub.(1-x)).sub.2 O.sub.3 buffer layer over the substrate, wherein 0laminate article can include a layer of YBCO over the (RE.sup.1.sub.x RE.sup.2.sub.(1-x)).sub.2 O.sub.3 buffer layer. A layer of CeO.sub.2 between the YBCO layer and the (RE.sup.1.sub.x RE.sup.2.sub.(1-x)).sub.2 O.sub.3 buffer can also be include. Further included can be a layer of YSZ between the CeO.sub.2 layer and the (R.sup.1.sub.x RE.sup.2.sub.(1-x)).sub.2 O.sub.3 buffer layer. The substrate can be a biaxially textured metal, such as nickel. A method of forming the laminate article is also disclosed.

  1. Superconductivity in Ca-doped graphene laminates.

    PubMed

    Chapman, J; Su, Y; Howard, C A; Kundys, D; Grigorenko, A N; Guinea, F; Geim, A K; Grigorieva, I V; Nair, R R

    2016-03-16

    Despite graphene's long list of exceptional electronic properties and many theoretical predictions regarding the possibility of superconductivity in graphene, its direct and unambiguous experimental observation has not been achieved. We searched for superconductivity in weakly interacting, metal decorated graphene crystals assembled into so-called graphene laminates, consisting of well separated and electronically decoupled graphene crystallites. We report robust superconductivity in all Ca-doped graphene laminates. They become superconducting at temperatures (Tc) between ≈4 and ≈6 K, with Tc's strongly dependent on the confinement of the Ca layer and the induced charge carrier concentration in graphene. We find that Ca is the only dopant that induces superconductivity in graphene laminates above 1.8 K among several dopants used in our experiments, such as potassium, caesium and lithium. By revealing the tunability of the superconducting response through doping and confinement of the metal layer, our work shows that achieving superconductivity in free-standing, metal decorated monolayer graphene is conditional on an optimum confinement of the metal layer and sufficient doping, thereby bringing its experimental realization within grasp.

  2. Vascular perfusion in horses with chronic laminitis.

    PubMed

    Hood, D M; Grosenbaugh, D A; Slater, M R

    1994-05-01

    Vascular perfusion casts were used to define and characterise the macroscopic perfusion defects present in the distal digit of 11 horses affected by chronic laminitis. Five clinically normal horses were used as controls. Based on clinical history and clinical status, horses with chronic laminitis were classified as being potentially treatable or clinically refractory. Eleven macroscopic vascular defects were noted in the casts from horses with laminitis. Four types of lesions were identified in the submural laminar circulation, 3 in the coronary bed and 4 were associated with the solar circulation. Multiple defects were present and a definite trend was noted for the perfusion defects to be worse in the casts of clinically refractory subjects than in those considered treatable. This information suggests that evaluation of circulatory status should add significantly to the ability to separate treatable from clinically refractory patients. Results also indicated that ventral displacement of the third phalanx (sinkers) and compression of the solar vasculature are more prevalent than is presently thought.

  3. Eastern Guaymas Basin: laminated but not anoxic

    NASA Astrophysics Data System (ADS)

    Cheshire, H.; Thurow, J. W.

    2009-12-01

    Site MD02-2513, eastern Guaymas Basin, challenges the accepted hypothesis about the circumstances and sequence of events leading to the occurrence of annually laminated (varved) sediment. By convention, under a climate regime of strong seasonal contrast, high productivity in a restricted basin enhances a preformed OMZ and inhibits bioturbation, resulting in the preservation of varves. The comparison of the high-resolution log of the sediment fabric and continuous XRF data from Core MD02-2513 reveal that laminated intervals over the last ~90 ka BP are characterised by an increase of oxygenation above background levels and are accompanied by lows of terrestrial and biogenic flux indicating that the preserving factor is rapid burial rather than high sedimentation rate (ie high seasonality and low rainfall). Intervals of diffuse/discontinuous laminations are times of high terrestrial and biogenic flux indicating comparatively low seasonality and high rainfall. Brief periods of relatively high anoxic conditions occur within homogeneous intervals and are accompanied by exceptionally high biogenic flux, indicating there was no seasonal contrast. Long homogenous intervals occur during the last glacial due to lowered sea level and the consequent removal of the upwelling cell offshore. The changes in Guaymas Basin seasonality are most probably caused by alterations to the migration pattern of the North Pacific Subtropical High due to the growth and decay of the North American ice sheets.

  4. Superconductivity in Ca-doped graphene laminates

    NASA Astrophysics Data System (ADS)

    Chapman, J.; Su, Y.; Howard, C. A.; Kundys, D.; Grigorenko, A. N.; Guinea, F.; Geim, A. K.; Grigorieva, I. V.; Nair, R. R.

    2016-03-01

    Despite graphene’s long list of exceptional electronic properties and many theoretical predictions regarding the possibility of superconductivity in graphene, its direct and unambiguous experimental observation has not been achieved. We searched for superconductivity in weakly interacting, metal decorated graphene crystals assembled into so-called graphene laminates, consisting of well separated and electronically decoupled graphene crystallites. We report robust superconductivity in all Ca-doped graphene laminates. They become superconducting at temperatures (Tc) between ≈4 and ≈6 K, with Tc’s strongly dependent on the confinement of the Ca layer and the induced charge carrier concentration in graphene. We find that Ca is the only dopant that induces superconductivity in graphene laminates above 1.8 K among several dopants used in our experiments, such as potassium, caesium and lithium. By revealing the tunability of the superconducting response through doping and confinement of the metal layer, our work shows that achieving superconductivity in free-standing, metal decorated monolayer graphene is conditional on an optimum confinement of the metal layer and sufficient doping, thereby bringing its experimental realization within grasp.

  5. Vibration analysis of bimodulus laminated cylindrical panels

    NASA Astrophysics Data System (ADS)

    Khan, K.; Patel, B. P.; Nath, Y.

    2009-03-01

    This paper deals with the flexural vibration behavior of bimodular laminated composite cylindrical panels with various boundary conditions. The formulation is based on first order shear deformation theory and Bert's constitutive model. The governing equations are derived using finite element method and Lagrange's equation of motion. An iterative eigenvalue approach is employed to obtain the positive and negative half cycle free vibration frequencies and corresponding mode shapes. A detailed parametric study is carried out to study the influences of thickness ratio, aspect ratio, lamination scheme, edge conditions and bimodularity ratio on the free vibration characteristics of bimodulus angle- and cross-ply composite laminated cylindrical panels. It is interesting to observe that there is a significant difference between the frequencies of positive and negative half cycles depending on the panel parameters. Through the thickness distribution of modal stresses for positive half cycle is significantly different from that for negative half cycle unlike unimodular case wherein the stresses at a particular location in negative half cycle would be of same magnitude but of opposite sign of those corresponding to positive half cycle. Finally, the effect of bimodularity on the steady state response versus forcing frequency relation is studied for a typical case.

  6. The prevalence of endocrinopathic laminitis among horses presented for laminitis at a first-opinion/referral equine hospital.

    PubMed

    Karikoski, N P; Horn, I; McGowan, T W; McGowan, C M

    2011-10-01

    Endocrinopathic causes of laminitis may be a common underlying causative pathogenesis in first-opinion or field cases presenting with laminitis, as opposed to laminitis produced in inflammatory research models. This study aimed to determine whether evidence of an underlying endocrinopathy was present in horses presented for laminitis to a first-opinion/referral veterinary teaching hospital. A second aim was to compare the signalment of horses and ponies with laminitis with the equine hospital population during the same period. All horses presenting for laminitis at Helsinki University Equine Teaching Hospital, Finland, over a 16-month period were examined for an underlying endocrinopathy. Horses presenting for laminitis were compared with the hospitalized population over the same period. There were 36 horses presented for laminitis, and evidence of endocrinopathy was present in 89%. Of the horses showing an underlying endocrinopathy, one-third had a diagnosis of pituitary pars intermedia dysfunction, and two-thirds showed basal hyperinsulinemia indicative of insulin resistance, without evidence of hirsutism. Phenotypic indicators of obesity were present in 95% of horses with basal hyperinsulinemia without hirsutism. Compared with the hospital population during the same period, horses with laminitis associated with an underlying endocrinopathy were significantly older and more likely to be pony breeds. Our data support that endocrine testing should be performed on all cases of laminitis that do not have a clear inflammatory or gastrointestinal origin. Copyright © 2011 Elsevier Inc. All rights reserved.

  7. Lamin-B1 contributes to the proper timing of epicardial cell migration and function during embryonic heart development

    PubMed Central

    Tran, Joseph R.; Zheng, Xiaobin; Zheng, Yixian

    2016-01-01

    Lamin proteins form a meshwork beneath the nuclear envelope and contribute to many different cellular processes. Mutations in lamins cause defective organogenesis in mouse models and human diseases that affect adipose tissue, brain, skeletal muscle, and the heart. In vitro cell culture studies have shown that lamins help maintain nuclear shape and facilitate cell migration. However, whether these defects contribute to improper tissue building in vivo requires further clarification. By studying the heart epicardium during embryogenesis, we show that Lb1-null epicardial cells exhibit in vivo and in vitro migratory delay. Transcriptome analyses of these cells suggest that Lb1 influences the expression of cell adhesion genes, which could affect cell migration during epicardium development. These epicardial defects are consistent with incomplete development of both vascular smooth muscle and compact myocardium at later developmental stages in Lb1-null embryos. Further, we found that Lb1-null epicardial cells have a delayed nuclear morphology change in vivo, suggesting that Lb1 facilitates morphological changes associated with migration. These findings suggest that Lb1 contributes to nuclear shape maintenance and migration of epicardial cells and highlights the use of these cells for in vitro and in vivo study of these classic cell biological phenomena. PMID:27798236

  8. Waved core lamination techniques on large and bulb hydroelectric machinery

    SciTech Connect

    Lehoczky, K.N. )

    1988-12-01

    Stator core lamination of large hydroelectric generators and motors and especially bulb type machines are often exposed to heat expansion forces causing severe damages. The temperature difference between the warm core lamination and the relatively colder stator frame, creates both radial and tangential stresses in these components. The consequence of these stresses depends on the axial prestressing of lamination and the internal design of the core. An overriding of some critical combination of these stresses may result in buckling of lamination, fracture of core tensioning and supporting components or cracks in the stator frame. The present waved lamination technique reduces the tangential and radial spring constant of core and prevents the damages. The research performed through theoretical and experimental methods, indicates a fundamental change in the behavior of core. The waved lamination was successfully used in a large number of machines, providing the reliability and wide application range of the method.

  9. Improved damage tolerance of titanium by adhesive lamination

    NASA Technical Reports Server (NTRS)

    Johnson, W. S.

    1982-01-01

    Basic damage tolerance properties of Ti-6A1-4V titanium plate can be improved by laminating thin sheets of titanium with adhesives. Compact tension and center cracked tension specimens made from thick plate, thin sheet, and laminated plate (six plies of thin sheet) were tested. The fracture toughness of the laminated plate was 39 percent higher than the monolithic plate. The laminated plate's through the thickness crack growth rate was about 20 percent less than that of the monolithic plate. The damage tolerance life of the surface cracked laminate was 6 to over 15 times the life of a monolithic specimen. A simple method of predicting crack growth in a crack ply of a laminate is presented.

  10. Homogenized Creep Behavior of CFRP Laminates at High Temperature

    NASA Astrophysics Data System (ADS)

    Fukuta, Y.; Matsuda, T.; Kawai, M.

    In this study, creep behavior of a CFRP laminate subjected to a constant stress is analyzed based on the time-dependent homogenization theory developed by the present authors. The laminate is a unidirectional carbon fiber/epoxy laminate T800H/#3631 manufactured by Toray Industries, Inc. Two kinds of creep analyses are performed. First, 45° off-axis creep deformation of the laminate at high temperature (100°C) is analyzed with three kinds of creep stress levels, respectively. It is shown that the present theory accurately predicts macroscopic creep behavior of the unidirectional CFRP laminate observed in experiments. Then, high temperature creep deformations at a constant creep stress are simulated with seven kinds of off-axis angles, i.e., θ = 0°, 10°, 30°, 45°, 60°, 75°, 90°. It is shown that the laminate has marked in-plane anisotropy with respect to the creep behavior.

  11. Thermo-mechanical response predictions for metal matrix composite laminates

    NASA Technical Reports Server (NTRS)

    Aboudi, J.; Hidde, J. S.; Herakovich, C. T.

    1991-01-01

    An analytical micromechanical model is employed for prediction of the stress-strain response of metal matrix composite laminates subjected to thermomechanical loading. The predicted behavior of laminates is based upon knowledge of the thermomechanical response of the transversely isotropic, elastic fibers and the elastic-viscoplastic, work-hardening matrix. The method is applied to study the behavior of silicon carbide/titanium metal matrix composite laminates. The response of laminates is compared with that of unidirectional lamina. The results demonstrate the effect of cooling from a stress-free temperature and the mismatch of thermal and mechanical properties of the constituent phases on the laminate's subsequent mechanical response. Typical results are presented for a variety of laminates subjected to monotonic tension, monotonic shear and cyclic tensile/compressive loadings.

  12. Processing and mechanical characterization of alumina laminates

    NASA Astrophysics Data System (ADS)

    Montgomery, John K.

    2002-08-01

    Single-phase ceramics that combine property gradients or steps in monolithic bodies are sought as alternatives to ceramic composites made of dissimilar materials. This work describes novel processing methods to produce stepped-density (or laminated) alumina single-phase bodies that maintain their mechanical integrity. One arrangement consists of a stiff, dense bulk material with a thin, flaw tolerant, porous exterior layer. Another configuration consists of a lightweight, low-density bulk material with a thin, hard, wear resistant exterior layer. Alumina laminates with strong interfaces have been successfully produced in this work using two different direct-casting processes. Gelcasting is a useful near-net shape processing technique that has been combined with several techniques, such as reaction bonding of aluminum oxide and the use of starch as a fugative filler, to successfully produced stepped-density alumina laminates. The other direct casting process that has been developed in this work is thermoreversible gelcasting (TRG). This is a reversible gelation process that has been used to produce near-net shape dense ceramic bodies. Also, individual layers can be stacked together and heated to produce laminates. Bilayer laminate samples were produced with varied thickness of porous and dense layers. It was shown that due to the difference in modulus and hardness, transverse cracking is found upon Hertzian contact when the dense layer is on the exterior. In the opposite arrangement, compacted damage zones formed in the porous material and no damage occurred in the underlying dense layer. Flaw tolerant behavior of the porous exterior/dense underlayer was examined by measuring biaxial strength as a function of Vickers indentation load. It was found that the thinnest layer of porous material results in the greatest flaw tolerance. Also, higher strength was exhibited at large indentation loads when compared to dense monoliths. The calculated stresses on the surfaces

  13. Arteriographical and pathological changes in chronic laminitis in dairy cattle.

    PubMed

    Boosman, R; Nemeth, F; Gruys, E; Klarenbeek, A

    1989-07-01

    The arteriographic appearance of 76 bovine hind digits, obtained from a slaughterhouse, was related to the macroscopic signs of chronic laminitis in the digits. There were statistically significant correlations between the macroscopic and the arteriographic appearance of the claws. Subsequent histological examination of the radiographically abnormal arteries revealed features indicative of arteriosclerosis. The results of this study indicate that chronic laminitis develops following a subclinical attack of laminitis due to a continous hypoperfusion of the digit.

  14. Impact Damage Tolerance of a Carbon Fibre Composite Laminate.

    DTIC Science & Technology

    1984-05-01

    design of composite structures. 8 CONCLUSIONS These carbon fibre/ epoxy resin laminates are susceptible :: low e ;rt., - .. impact damage, especially...ROYAL AIRCRAFT ESTABLISHMENT0 Technical Report 84049 May 1984 GARTEUR/TP-007 IMPACT DAMAGE TOLERANCE OF A CARBON FIBRE COMPOSITE LAMINATE by DTIC G...007 Received for printing 3 May 1984 IMPACT DAMAGE TOLERANCE OF A CARBON FIBRE COMPOSITE LAMINATE by G. Dorey P. Sigety* K. Stellbrink** W. G. J. ’t

  15. Non-linear behavior of fiber composite laminates

    NASA Technical Reports Server (NTRS)

    Hashin, Z.; Bagchi, D.; Rosen, B. W.

    1974-01-01

    The non-linear behavior of fiber composite laminates which results from lamina non-linear characteristics was examined. The analysis uses a Ramberg-Osgood representation of the lamina transverse and shear stress strain curves in conjunction with deformation theory to describe the resultant laminate non-linear behavior. A laminate having an arbitrary number of oriented layers and subjected to a general state of membrane stress was treated. Parametric results and comparison with experimental data and prior theoretical results are presented.

  16. Methods for Preparing Nanoparticle-Containing Thermoplastic Composite Laminates

    NASA Technical Reports Server (NTRS)

    Gruber, Mark B. (Inventor); Jensen, Brian J. (Inventor); Cano, Roberto J. (Inventor)

    2016-01-01

    High quality thermoplastic composites and composite laminates containing nanoparticles and/or nanofibers, and methods of producing such composites and laminates are disclosed. The composites comprise a thermoplastic polymer and a plurality of nanoparticles, and may include a fibrous structural reinforcement. The composite laminates are formed from a plurality of nanoparticle-containing composite layers and may be fused to one another via an automated process.

  17. Fracture behavior of thick, laminated graphite/epoxy composites

    NASA Technical Reports Server (NTRS)

    Harris, C. E.; Morris, D. H.

    1984-01-01

    The effect of laminate thickness on the fracture behavior of laminated graphite epoxy (T300/5208) composites was studied. The predominantly experimental research program included the study of the 0/+ or - 45/90 sub ns and 0/90 sub ns laminates with thickness of 8, 32, 64, 96 and 120 plies and the 0/+ or - 45 sub ns laminate with thickness of 6, 30, 60, 90 and 120 plies. The research concentrated on the measurement of fracture toughness utilizing the center-cracked tension, compact tension and three point bend specimen configurations. The development of subcritical damage at the crack tip was studied nondestructively using enhanced X-ray radiography and destructively using the laminate deply technique. The test results showed fracture toughness to be a function of laminate thickness. The fracture toughness of the 0 + or - 45/90 sub ns and 0/90 sub ns laminates decreased with increasing thickness and asymptotically approached lower bound values of 30 ksi square root of in. (1043 MPa square root of mm and 25 ksi square root of in (869 MPa square root of mm respectively. In contrast to the other two laminates, the fracture toughness of the 0/+ or - 45 sub ns laminate increased sharply with increasing thickness but reached an upper plateau value of 40 ksi square root of in (1390 MPa square root of mm) at 30 plies. Fracture toughness was independent of crack size for both thin and thick laminates for all three laminate types except for the 0/90 sub 2s laminate which spilt extensively. The center cracked tension, three point bend and compact tension specimens gave comparable results.

  18. Energy Saving Glass Lamination via Selective Radio-Frequency Heating

    SciTech Connect

    Shulman, Holly S.; Allan, Shawn M.

    2009-11-11

    This Inventions and Innovations program supported the technical and commercial research and development needed to elevate Ceralink's energy saving process for flat glass lamination from bench scale to a self-supporting technology with significant potential for growth. Radio-frequency heating was any un-explored option for laminating glass prior to this program. With significant commercial success through time and energy savings in the wood, paper, and plastics industries, RF heating was found to have significant promise for the energy intensive glass lamination industry. A major technical goal of the program was to demonstrate RF lamination across a wide range of laminate sizes and materials. This was successfully accomplished, dispelling many skeptics' concerns about the abilities of the technology. Ceralink laminated panels up to 2 ft x 3 ft, with four sets processed simultaneously, in a 3 minute cycle. All major categories of interlayer materials were found to work with RF lamination. In addition to laminating glass, other materials including photovoltaic silicon solar cells, light emitting diodes, metallized glass, plastics (acrylic and polycarbonate), and ceramics (alumina) were found compatible with the RF process. This opens up a wide range of commercial opportunities beyond the initially targeted automotive industry. The dramatic energy savings reported for RF lamination at the bench scale were found to be maintained through the scale up of the process. Even at 2 ft x 3 ft panel sizes, energy savings are estimated to be at least 90% compared to autoclaving or vacuum lamination. With targeted promotion through conference presentations, press releases and internet presence, RF lamination has gained significant attention, drawing large audiences at American Ceramic Society meetings. The commercialization success of the project includes the establishment of a revenue-generating business model for providing process development and demonstrations for potential RF

  19. Energy Saving Glass Lamination via Selective Radio Frequency Heating

    SciTech Connect

    Allan, Shawn M.

    2012-02-27

    This project focused on advancing radio-frequency (RF) lamination technology closer to commercial implementation, in order to reduce the energy intensity of glass lamination by up to 90%. Lamination comprises a wide range of products including autoglass, architectural safety and innovative design glass, transparent armor (e.g. bullet proof glass), smart glass, mirrors, and encapsulation of photovoltaics. Lamination is also the fastest growing segment of glass manufacturing, with photovoltaics, architectural needs, and an anticipated transition to laminated side windows in vehicles. The state-of-the-art for glass lamination is to use autoclaves, which apply heat and uniform gas pressure to bond the laminates over the course of 1 to 18 hours. Laminates consist of layers of glass or other materials bonded with vinyl or urethane interlayers. In autoclaving, significant heat energy is lost heating the chamber, pressurized air, glass racks, and the glass. In RF lamination, the heat is generated directly in the vinyl interlayer, causing it to heat and melt quickly, in just 1 to 10 minutes, without significantly heating the glass or the equipment. The main purpose of this project was to provide evidence that low energy, rapid RF lamination quality met the same standards as conventionally autoclaved windows. The development of concepts for laminating curved glass with RF lamination was a major goal. Other primary goals included developing a stronger understanding of the lamination product markets described above, and to refine the potential benefits of commercial implementation. The scope of the project was to complete implementation concept studies in preparation for continuation into advanced development, pilot studies, and commercial implementation. The project consisted of 6 main tasks. The first dealt with lamination with poly-vinyl butyral (PVB) interlayers, which prior work had shown difficulties in achieving good quality laminates, working with Pilkington North

  20. Energy Saving Glass Lamination via Selective Radio Frequency Heating

    SciTech Connect

    Allan, Shawn M; Baranova, Inessa; Poley, Joseph; Reis, Henrique

    2012-02-27

    This project focused on advancing radio-frequency (RF) lamination technology closer to commercial implementation, in order to reduce the energy intensity of glass lamination by up to 90%. Lamination comprises a wide range of products including autoglass, architectural safety and innovative design glass, transparent armor (e.g. bullet proof glass), smart glass, mirrors, and encapsulation of photovoltaics. Lamination is also the fastest growing segment of glass manufacturing, with photovoltaics, architectural needs, and an anticipated transition to laminated side windows in vehicles. The state-of-the-art for glass lamination is to use autoclaves, which apply heat and uniform gas pressure to bond the laminates over the course of 1 to 18 hours. Laminates consist of layers of glass or other materials bonded with vinyl or urethane interlayers. In autoclaving, significant heat energy is lost heating the chamber, pressurized air, glass racks, and the glass. In RF lamination, the heat is generated directly in the vinyl interlayer, causing it to heat and melt quickly, in just 1 to 10 minutes, without significantly heating the glass or the equipment. The main purpose of this project was to provide evidence that low energy, rapid RF lamination quality met the same standards as conventionally autoclaved windows. The development of concepts for laminating curved glass with RF lamination was a major goal. Other primary goals included developing a stronger understanding of the lamination product markets described above, and to refine the potential benefits of commercial implementation. The scope of the project was to complete implementation concept studies in preparation for continuation into advanced development, pilot studies, and commercial implementation. The project consisted of 6 main tasks. The first dealt with lamination with poly-vinyl butyral (PVB) interlayers, which prior work had shown difficulties in achieving good quality laminates, working with Pilkington North

  1. Residual stress and crack propagation in laminated composites

    SciTech Connect

    Yttergren, R.M.F.; Zeng, K.; Rowcliffe, D.J.

    1994-12-31

    Residual stress distributions in several laminated ceramic composites were measured by an indentation technique. The material included alumina-zirconia laminated composites, containing strong interfaces, and alumina-porcelain laminated composites with both weak and strong interfaces. The residual stress in these material originates from the mismatch of the thermal properties, differences in elastic properties, and different shrinkage of the laminates during sintering. An experimental technique is presented which gives a direct view of the residual stress state in the materials. Values of residual tensile stress are presented as a function of position relative to the interface in each material.

  2. Extrinsic fracture mechanisms in two laminated metal composites

    SciTech Connect

    Lesuer, D.; Syn, C.; Riddle, R.; Sherby, O.

    1994-11-29

    The crack growth behavior and fracture toughness of two laminated metal composites (6090/SiC/25p laminated with 5182 and ultrahigh-carbon steel laminated with brass) have been studied in both ``crack arrester`` and ``crack divider`` orientations. The mechanisms of crack growth were analyzed and extrinsic toughening mechanisms were found to contribute significantly to the toughness. The influence of laminate architecture (layer thickness and component volume function), component material properties and residual stress on these mechanisms and the resulting crack growth resistance are discussed.

  3. Impact damage resistance of thin stitched carbon/epoxy laminates

    NASA Astrophysics Data System (ADS)

    Francesconi, L.; Aymerich, F.

    2015-07-01

    The study examines the influence of through-thickness stitching on the damage response of thin cross-ply carbon/epoxy laminates subjected to low-velocity impacts. Instrumented impact tests were carried out on unstitched and polyethylene stitched laminates and the resulting damage was assessed in detail by X-radiography analyses. The results of the observations carried out during the experimental analyses are illustrated and discussed to identify the mechanical role played by through-thickness reinforcement and to highlight the influence of the laminate layup on the impact resistance of stitched laminates.

  4. A simple and high-throughput method to assess maturation status of bovine oocytes: comparison of anti-lamin A/C-DAPI with an aceto-orcein staining technique.

    PubMed

    Prentice-Biensch, J R; Singh, J; Alfoteisy, B; Anzar, M

    2012-10-15

    A precise, accurate, nonambiguous and high-throughput method is required to assess nuclear maturation of mammalian oocytes. The objectives of this study were to compare the efficiency and ease of use of a simplified fluorescence imaging (anti-lamin A/C and 4',6-diamidino-2-phenylindole [DAPI]) technique to the existing technique (aceto-orcein staining) for the evaluation of nuclear maturation of bovine oocytes, and to determine the kinetics of bovine oocyte maturation using an anti-lamin A/C-DAPI technique. In Experiment 1, oocytes were matured in vitro and stained with aceto-orcein and anti-lamin A/C-DAPI staining techniques. The proportions of oocytes lost during procedures and those that could not be classified (because of ambiguous morphology) during evaluation were lower (P < 0.0001) in oocytes stained with anti-lamin A/C-DAPI (9% and 2%) than those stained with aceto-orcein (31% and 13%), respectively. Anti-lamin A/C-DAPI was a quick procedure which could be completed within 7 h after completion of the maturation (compared with > 24 h for the aceto-orcein method). Furthermore, > 200 oocytes could be stained in one batch with anti-lamin A/C-DAPI technique. In Experiment 2, nuclear maturation kinetics of bovine oocytes at various time intervals (0, 6, 12, and 22 h) during in vitro maturation (IVM) was evaluated using the anti-lamin A/C-DAPI technique. Germinal vesicle, germinal vesicle breakdown, metaphase I, and metaphase II oocytes were predominant at 0, 6, 12, and 22 h of IVM, respectively. We concluded that the anti-lamin A/C-DAPI was an efficient and simple technique for nonambiguous evaluation of nuclear maturation status of large numbers of oocytes in a short interval. Copyright © 2012 Elsevier Inc. All rights reserved.

  5. Fission yeast Lem2 and Man1 perform fundamental functions of the animal cell nuclear lamina.

    PubMed

    Gonzalez, Yanira; Saito, Akira; Sazer, Shelley

    2012-01-01

    In animal cells the nuclear lamina, which consists of lamins and lamin-associated proteins, serves several functions: it provides a structural scaffold for the nuclear envelope and tethers proteins and heterochromatin to the nuclear periphery. In yeast, proteins and large heterochromatic domains including telomeres are also peripherally localized, but there is no evidence that yeast have lamins or a fibrous nuclear envelope scaffold. Nonetheless, we found that the Lem2 and Man1 proteins of the fission yeast Schizosaccharomyces pombe, evolutionarily distant relatives of the Lap2/Emerin/Man1 (LEM) sub-family of animal cell lamin-associated proteins, perform fundamental functions of the animal cell lamina. These integral inner nuclear membrane localized proteins, with nuclear localized DNA binding Helix-Extension-Helix (HEH) domains, impact nuclear envelope structure and integrity, are essential for the enrichment of telomeres at the nuclear periphery and by means of their HEH domains anchor chromatin, most likely transcriptionally repressed heterochromatin, to the nuclear periphery. These data indicate that the core functions of the nuclear lamina are conserved between fungi and animal cells and can be performed in fission yeast, without lamins or other intermediate filament proteins.

  6. Fission yeast Lem2 and Man1 perform fundamental functions of the animal cell nuclear lamina

    PubMed Central

    Gonzalez, Yanira; Saito, Akira; Sazer, Shelley

    2012-01-01

    In animal cells the nuclear lamina, which consists of lamins and lamin-associated proteins, serves several functions: it provides a structural scaffold for the nuclear envelope and tethers proteins and heterochromatin to the nuclear periphery. In yeast, proteins and large heterochromatic domains including telomeres are also peripherally localized, but there is no evidence that yeast have lamins or a fibrous nuclear envelope scaffold. Nonetheless, we found that the Lem2 and Man1 proteins of the fission yeast Schizosaccharomyces pombe, evolutionarily distant relatives of the Lap2/Emerin/Man1 (LEM) sub-family of animal cell lamin-associated proteins, perform fundamental functions of the animal cell lamina. These integral inner nuclear membrane localized proteins, with nuclear localized DNA binding Helix-Extension-Helix (HEH) domains, impact nuclear envelope structure and integrity, are essential for the enrichment of telomeres at the nuclear periphery and by means of their HEH domains anchor chromatin, most likely transcriptionally repressed heterochromatin, to the nuclear periphery. These data indicate that the core functions of the nuclear lamina are conserved between fungi and animal cells and can be performed in fission yeast, without lamins or other intermediate filament proteins. PMID:22540024

  7. Accelerated Telomere Shortening and Replicative Senescence in Human Fibroblasts Overexpressing Mutant and Wild Type Lamin A

    PubMed Central

    Huang, Shurong; Risques, Rosa Ana; Martin, George M.; Rabinovitch, Peter S.; Oshima, Junko

    2008-01-01

    LMNA mutations are responsible for a variety of genetic disorders, including muscular dystrophy, lipodystrophy, and certain progeroid syndromes, notably Hutchinson-Gilford Progeria. Although a number of clinical features of these disorders are suggestive of accelerated aging, it is not known whether cells derived from these patients exhibit cellular phenotypes associated with accelerated aging. We examined a series of isogenic skin fibroblast lines transfected with LMNA constructs bearing known pathogenic point mutations or deletion mutations found in progeroid syndromes. Fibroblasts overexpressing mutant lamin A exhibited accelerated rates of loss of telomeres and shortened replicative lifespans, in addition to abnormal nuclear morphology. To our surprise, these abnormalities were also observed in lines overexpressing wild-type lamin A. Copy number variants are common in human populations; those involving LMNA, whether arising meiotically or mitotically, might lead to progeroid phenotypes. In an initial pilot study of 23 progeroid cases without detectible WRN or LMNA mutations, however, no cases of altered LMNA copy number were detected. Nevertheless, our findings raise a hypothesis that changes in lamina organization may cause accelerated telomere attrition, with different kinetics for overexpession of wild-type and mutant lamin A, which leads to rapid replicative senescence and progroid phenotypes. PMID:17870066

  8. Strength and Stiffness of Small Glued-Laminated Beams with Different Qualities of Tension Laminations.

    DTIC Science & Technology

    1981-05-01

    difference two of the Li DF-L tension lamina- other test material in this study and between the average of the unad - tions were chosen for that reason...MOE data, adjusted to a 12 per- of tension lamination had a signifi- should be combined to determine the cent moisture content, and the unad - cant

  9. Causes and consequences of nuclear envelope alterations in tumour progression.

    PubMed

    Bell, Emily S; Lammerding, Jan

    2016-11-01

    Morphological changes in the size and shape of the nucleus are highly prevalent in cancer, but the underlying molecular mechanisms and the functional relevance remain poorly understood. Nuclear envelope proteins, which can modulate nuclear shape and organization, have emerged as key components in a variety of signalling pathways long implicated in tumourigenesis and metastasis. The expression of nuclear envelope proteins is altered in many cancers, and changes in levels of nuclear envelope proteins lamins A and C are associated with poor prognosis in multiple human cancers. In this review we highlight the role of the nuclear envelope in different processes important for tumour initiation and cancer progression, with a focus on lamins A and C. Lamin A/C controls many cellular processes with key roles in cancer, including cell invasion, stemness, genomic stability, signal transduction, transcriptional regulation, and resistance to mechanical stress. In addition, we discuss potential mechanisms mediating the changes in lamin levels observed in many cancers. A better understanding of cause-and-effect relationships between lamin expression and tumour progression could reveal important mechanisms for coordinated regulation of oncogenic processes, and indicate therapeutic vulnerabilities that could be exploited for improved patient outcome.

  10. Systematic Analysis of the Functional Relevance of Nuclear Structure and Mechanics in Breast Cancer Progression

    DTIC Science & Technology

    2013-07-01

    ANSI Std. Z39.18 Systematic Analysis of the Functional Relevance of Nuclear Structure and Mechanics in Breast Cancer Progression Jan Lammerding... analysis of the functional consequences of changes in the expression of lamins (A, B1, B2, and C) and lamin B receptor on nuclear morphology and...enhanced passage), proliferation, and epithelial-to- mesenchymal transition (EMT). In addition, we proposed to conduct an analysis of samples

  11. Neuropathic changes in equine laminitis pain.

    PubMed

    Jones, Emma; Viñuela-Fernandez, Ignacio; Eager, Rachel A; Delaney, Ada; Anderson, Heather; Patel, Anisha; Robertson, Darren C; Allchorne, Andrew; Sirinathsinghji, Eva C; Milne, Elspeth M; MacIntyre, Neil; Shaw, Darren J; Waran, Natalie K; Mayhew, Joe; Fleetwood-Walker, Susan M

    2007-12-05

    Laminitis is a common debilitating disease in horses that involves painful disruption of the lamellar dermo-epidermal junction within the hoof. This condition is often refractory to conventional anti-inflammatory analgesia and results in unremitting pain, which in severe cases requires euthanasia. The mechanisms underlying pain in laminitis were investigated using quantification of behavioural pain indicators in conjunction with histological studies of peripheral nerves innervating the hoof. Laminitic horses displayed consistently altered or abnormal behaviours such as increased forelimb lifting and an increased proportion of time spent at the back of the box compared to normal horses. Electron micrographic analysis of the digital nerve of laminitic horses showed peripheral nerve morphology to be abnormal, as well as having reduced numbers of unmyelinated (43.2%) and myelinated fibers (34.6%) compared to normal horses. Sensory nerve cell bodies innervating the hoof, in cervical, C8 dorsal root ganglia (DRG), showed an upregulated expression of the neuronal injury marker, activating transcription factor-3 (ATF3) in both large NF-200-immunopositive neurons and small neurons that were either peripherin- or IB4-positive. A significantly increased expression of neuropeptide Y (NPY) was also observed in myelinated afferent neurons. These changes are similar to those reported in other neuropathic pain states and were not observed in the C4 DRG of laminitic horses, which is not associated with innervation of the forelimb. This study provides novel evidence for a neuropathic component to the chronic pain state associated with equine laminitis, indicating that anti-neuropathic analgesic treatment may well have a role in the management of this condition.

  12. Expression of endothelin in equine laminitis.

    PubMed

    Katwa, L C; Johnson, P J; Ganjam, V K; Kreeger, J M; Messer, N T

    1999-05-01

    Biosynthesis of endothelin-1 (ET-1), the most potent endogenous vasoconstrictor yet identified, is increased following myocardial infarction (MI) in man. Pathological events which occur in the connective tissues of the equine hoof during laminitis are similar in some respects, to changes occurring in the myocardial connective tissues following MI in man. The objective of this study was to determine whether ET-1 expression in connective tissues obtained from the hoof of laminitic horses is increased compared with tissues obtained from healthy horses. Expression of ET-1 in connective tissues of the equine hoof was measured following tissue extraction from 3 groups of horses: horses in which acute laminitis had been induced by the administration of starch; chronically foundered horses; nonlaminitic horses. The concentration of ET-1 in laminar connective tissues obtained from all laminitic horses (1573.0 +/- 392.8 pg/g of tissue; n = 10) was increased when compared with tissues obtained from nonlaminitic horses (392.5 +/- 117.4 pg/g of tissue; n = 5) (P<0.05). The concentration of ET-1 in laminar connective tissues obtained from the experimentally induced, acute laminitic horses (1043.6 +/- 254.4 pg/g of tissue; n = 7) and from the spontaneously affected, chronic laminitic horses (2808.3 +/- 878.6 pg/g of tissue; n = 3) was increased compared with the control group (P<0.05, P<0.01, respectively). The concentration of ET-1 in laminar connective tissues obtained from the chronic laminitic horses was greater than that of the experimentally induced, acute laminitic group (P<0.05). It is suggested that the data provide a strong argument that increased ET-1 expression in the connective tissues of the equine hoof represent a potentially important and hitherto unrecognised component of the pathophysiology of equine laminitis. Further studies are needed to determine whether inhibitors of ET-1 converting enzyme or antagonists of ET-1 receptors might be useful in the treatment

  13. Micromechanics of composite laminate compression failures

    NASA Technical Reports Server (NTRS)

    Guynn, E. Gail; Bradley, Walter L.

    1988-01-01

    The purpose of this annual progress report is to summarize the work effort and results accomplished from July 1987 through July 1988 on NASA Research Grant NAG1-659 entitled Micromechanics of Composite Laminate Compressive Failure. The report contains: (1) the objective of the proposed research, (2) the summary of accomplishments, (3) a more extensive review of compression literature, (4) the planned material (and corresponding properties) received to date, (5) the results for three possible specimen geometries, experimental procedures planned, and current status of the experiments, and (6) the work planned for the next contract year.

  14. Tailoring Laminates For Protection Against Projectiles

    NASA Technical Reports Server (NTRS)

    Gassner, John J.; Boyce, Joseph S.; Smirlock, Martin E.; Evans, David A.

    1992-01-01

    Fiber/matrix composite laminates developed to protect military land vehicles against projectiles and spacecraft against impacts by micrometeroids. Although types, sizes, and velocities of expected incident objects differ between terrestrial and outer-space cases, general protection problems and solutions exhibit some common features. Configurations of fibers and matrices optimized with respect to protection, bulk, and weight, to obtain shield that breaks rapidly moving incident object into harmless smaller, more-slowly-moving pieces, containing debris, vaporizes debris, and/or otherwise absorbs kinetic energy of object to prevent harm to vehicle and occupants.

  15. Tailoring Laminates For Protection Against Projectiles

    NASA Technical Reports Server (NTRS)

    Gassner, John J.; Boyce, Joseph S.; Smirlock, Martin E.; Evans, David A.

    1992-01-01

    Fiber/matrix composite laminates developed to protect military land vehicles against projectiles and spacecraft against impacts by micrometeroids. Although types, sizes, and velocities of expected incident objects differ between terrestrial and outer-space cases, general protection problems and solutions exhibit some common features. Configurations of fibers and matrices optimized with respect to protection, bulk, and weight, to obtain shield that breaks rapidly moving incident object into harmless smaller, more-slowly-moving pieces, containing debris, vaporizes debris, and/or otherwise absorbs kinetic energy of object to prevent harm to vehicle and occupants.

  16. Fracture modes in notched angleplied composite laminates

    NASA Technical Reports Server (NTRS)

    Irvine, T. B.; Ginty, C. A.

    1984-01-01

    The Composite Durability Structural Analysis (CODSTRAN) computer code is used to determine composite fracture. Fracture modes in solid and notched, unidirectional and angleplied graphite/epoxy composites were determined by using CODSTRAN. Experimental verification included both nondestructive (ultrasonic C-Scanning) and destructive (scanning electron microscopy) techniques. The fracture modes were found to be a function of ply orientations and whether the composite is notched or unnotched. Delaminations caused by stress concentrations around notch tips were also determined. Results indicate that the composite mechanics, structural analysis, laminate analysis, and fracture criteria modules embedded in CODSTRAN are valid for determining composite fracture modes.

  17. Laminated anisotropic reinforced plastic plates and shells

    NASA Technical Reports Server (NTRS)

    Korolev, V. I.

    1981-01-01

    Basic technical theories and engineering calculation equations for anisotropic plates and shells made of rigid reinforced plastics, mainly laminated fiberglass, are presented and discussed. Solutions are given for many problems of design of structural plates and shells, including curved sections and tanks, as well as two chapters on selection of the optimum materials, are given. Accounting for interlayer shearing and transverse separation, which are new engineering properties, are discussed. Application of the results obtained to thin three ply plates and shells wth a light elastic filler is presented and discussed.

  18. Adhesive characterization in prestressed piezoelectric laminates

    NASA Astrophysics Data System (ADS)

    Hodges, Charles A.; Mossi, Karla M.; Scott, Lisa A.

    2003-08-01

    Pre-stressed piezoelectric laminates, consisting of one or more metal layers and a piezoelectric material bonded together with an adhesive, have been widely studied over the past few years, both numerically and experimentally. Most of the current research has concentrated on the effect of the metal layers, types and geometry, along with variations in the active layer of the laminate. Historically, the adhesive layer has been neglected as a contributing factor in the overall performance of the final device. This paper attempts to address the effect of the adhesive line thickness and its influence on the performance of pre-stressed piezoelectric laminates under specific boundary conditions. All laminates tested were constructed with the following lay-up: 0.354 mm thick stainless steel, adhesive, 0.381 mm PZT ceramic, adhesive, and a 0.0254 mm aluminum layer. The devices having an adhesive line thickness of 0.169 mm were classified as group A, and group B were the devices with an adhesive line thickness of 0.036 mm. The adhesive line thickness for group A was approximately 21% more than the line thickness of group B. The devices were tested in a simply supported, free-free condition under a series of loads at a constant frequency of 5 Hz over a voltage range from 400 to 800 Volts peak-to-peak. Displacement was measured using loads of 25, 50, 75, 100, and 200 grams for each actuator. The data from each group was averaged and compared. The results showed group B generated more displacement at the same "arm weight" applied as compared to group A. However, only three samples for group B were measured since the rest of the samples failed during testing. Failure of the devices of group B may be due to the ultimate stress of the devices and their ability to lift a load under those conditions. The study demonstrated that adhesive layer thickness, along with the manufacturing process, has to be taken into account when developing an application that requires load

  19. Strength of Bolted Joints in Laminated Composites

    DTIC Science & Technology

    1984-03-01

    considered (c13ao23=033a0). Under these conditions, in the absence of body forces, the condition of force equilibriui can be expressed as E18 ] (1) Bo2...Conference), ASTM STP 617, 1977, pp 229-242. 14. I.M. Daniel, R.E. Rowlands, and J.B. Whiteside, "Effects of Material and Stacking Sequence on the...Whitney, "Uniaxial Failure of Composite Laminates Containing Stress Concentrations,"Fracture Mechanics of Compo-ites, ASTM STP 593, 1975, pp. 117-142

  20. Buckling of angle-ply laminated circular cylindrical shells

    NASA Technical Reports Server (NTRS)

    Hirano, Y.

    1979-01-01

    This note presents closed-form solutions for axisymmetrical and axially unsymmetrical buckling of angle-ply laminated circular cylindrical shells under axial compression. The axisymmetrical and axially unsymmetrical buckling stress are found to be different from each other, and the best lamination angles which give the highest buckling stress are obtained.

  1. Mechanical performance of cellulose nanofibril film-wood flake laminate

    Treesearch

    Jen-Chieh Liu; Robert J. Moon; Alan Rudie; Jeffrey P. Youngblood

    2014-01-01

    Homogeneous and transparent CNF films, fabricated from the (2,2,6,6- tetramethylpiperidin-1-yl) oxyl (TEMPO)-modified CNF suspension, were laminated onto wood flakes (WF) based on phenol-formaldehyde (PF) resin and the reinforcement potential of the material has been investigated. The focus was on the influence of CNF film lamination, relative humidity (RH), heat...

  2. Maleated polypropylene film and wood fiber handsheet laminates

    Treesearch

    Sangyeob Lee; Todd F. Shupe; Leslie H. Groom; Chung Y. Hse

    2008-01-01

    The grafting effect of maleic anhydride (MA) as an interfacial bonding agent and its influence on the tensile strength properties of thermomechanical pulp handsheet-isotactic polypropylene (iPP) film laminates was studied. For the MA treated with benzoyl peroxide (BPO) as an initiator, tensile strength properties increased 76% with PP film over untreated laminates. The...

  3. Hole-thru-laminate mounting supports for photovoltaic modules

    DOEpatents

    Wexler, Jason; Botkin, Jonathan; Culligan, Matthew; Detrick, Adam

    2015-02-17

    A mounting support for a photovoltaic module is described. The mounting support includes a pedestal having a surface adaptable to receive a flat side of a photovoltaic module laminate. A hole is disposed in the pedestal, the hole adaptable to receive a bolt or a pin used to couple the pedestal to the flat side of the photovoltaic module laminate.

  4. Processing of thermoset prepreg laminate via exposure to microwave radiation

    SciTech Connect

    Paulauskas, F.L.; Meek, T.T.

    1994-06-01

    Studies of microwave-assisted curing of neat resins (epoxy) and unidirectional glass and carbon fiber laminates have shown that a substantial reduction in the curing time was obtained. This may be explained by the penetration of microwave energy directly and throughout the laminate with enhancement of the kinetics of the chemical reaction. Results of this work indicate that the microwave assisted curing of glass fiber laminates also shows a substantial reduction of the required curing time. Microwave radiation of 2.45 GHz has been demonstrated to be an acceptable method to cure unidirectional carbon fiber laminates. Also, effective curing of crossply (0/90) laminates through this method was observed when proper rotation of the parts accompanied the curing process. This is in accordance with previous work. Multidirectional carbon fiber/epoxy laminates demonstrate a lack of coupling during the curing process. A direct curing of these laminates was not possible by microwave radiation with the experimental approach used, in agreement with previous work. Nevertheless, a moderate reduction in the curing time of these thin laminates was observed due to hybrid curing.

  5. The North American Product Standard for Cross-Laminated Timber

    Treesearch

    Borjen Yeh; Sylvain Gagnon; Tom Williamson; Ciprian Pirvu; Conroy Lum; Dave Kretschmann

    2012-01-01

    Cross-laminated timber (CLT) is a prefabricated solid engineered wood product made of at least three orthogonally bonded layers of solid-sawn lumber or structural composite lumber that are laminated by gluing of longitudinal and transverse layers with structural adhesives to form a solid rectangular-shaped, straight, and plane timber intended for roof, floor, or wall...

  6. Lateral testing of glued laminated timber tudor arch

    Treesearch

    Douglas R. Rammer; Philip Line

    2016-01-01

    Glued laminated timber Tudor arches have been in wide use in the United States since the 1930s, but detailed knowledge related to seismic design in modern U.S. building codes is lacking. FEMA P-695 (P-695) is a methodology to determine seismic performance factors for a seismic force resisting system. A limited P-695 study for glued laminated timber arch structures...

  7. The pharmacologic basis for the treatment of endocrinopathic laminitis.

    PubMed

    Durham, Andy

    2010-08-01

    Although the treatment and management of laminitis in the horse requires a holistic and often multidisciplinary approach from the veterinarian, farrier, and nutritionist, this review focuses on pharmacologic interventions that might have prophylactic benefit, specifically in the horse with laminitis as a result of pituitary pars intermedia dysfunction and equine metabolic syndrome.

  8. Laminated sheet composites reinforced with modular filament sheet

    NASA Technical Reports Server (NTRS)

    Reece, O. Y.

    1968-01-01

    Aluminum and magnesium composite sheet laminates reinforced with low density, high strength modular filament sheets are produced by diffusion bonding and explosive bonding. Both processes are accomplished in normal atmosphere and require no special tooling or cleaning other than wire brushing the metal surfaces just prior to laminating.

  9. Fracture toughness and impact properties of laminated metal composites

    SciTech Connect

    Lesuer, D.R.; Riddle, R.A.; Gogolewski, R.P.; Syn, C.K.; Cunningham, B.J.

    1996-03-04

    Laminated metal composites consist of alternating metal (or metal matrix composite) layers bonded together. These materials can provide fracture toughness and impact properties superior to those of the component materials. These properties are a function of component material properties, laminate architecture (volume fraction, thickness) and interface properties. Properties are compared for seven lightweight materials.

  10. Chapter 2: Manufacturing Cross-laminated timber manufacturing

    Treesearch

    Borjen Yeh; Dave Kretschmann; Brad (Jianhe) Wang

    2013-01-01

    Cross-laminated timber ( CLT) is defined as a prefabricated solid engineered wood product made of at least three orthogonally bonded layers of solid-sawn lumber or structural composite lumber (SCL) that are laminated by gluing oflongitudinal and transverse layers with structural adhesives to form a solid rectangular-shaped, straight, and plane timber intended for roof...

  11. Better Thermal Insulation in Solar-Array Laminators

    NASA Technical Reports Server (NTRS)

    Burger, D. R.; Knox, J. F.

    1984-01-01

    Glass marbles improve temperature control. Modified vacuum laminator for photovoltaic solar arrays includes thermal insulation made of conventional glass marbles. Marbles serve as insulation for temperature control of lamination process at cure temperatures as high as 350 degrees F. Used to replace original insulation made of asbestos cement.

  12. Histopathology of oligofructose-induced acute laminitis in heifers.

    PubMed

    Thoefner, M B; Wattle, O; Pollitt, C C; French, K R; Nielsen, S S

    2005-08-01

    Histopathology of the dermo-epidermal junction in the lamellar region of front claws was examined in 6 dairy heifers given an alimentary oligofructose overload and compared with sections from a control group of 6 heifers. Four of the 6 heifers administered oligofructose developed clinical signs of acute laminitis before they were euthanized. Postmortem samples from front claws were processed for histology. Eleven histopathologic characteristics were selected from the existing literature and used in a blinded evaluation of sections. In total, 104 front claw samples, including 8 samples from 2 cows having spontaneously occurring acute laminitis, were evaluated histologically using hematoxylin and eosin as well as periodic acid-Schiff staining. The major morphological features associated with oligofructose-induced acute clinical laminitis were stretching of lamellae, dermal edema, hemorrhage, changes in basal cell morphology, presence of white blood cells in dermis, and signs of basement membrane detachment. Changes at the lamellar junction of claw tissue affected by oligofructose-induced clinical laminitis resembled tissue from the 2 cows suffering from spontaneous acute clinical laminitis, and generally were consistent with existing descriptions of laminitis histopathology. Important exceptions to existing descriptions in the literature were stretching of lamellae and basement membrane changes. Not previously described, we considered these early signs of acute laminitis. In conclusion, this study documents that oligofructose-induced clinical laminitis is associated with histopathological changes at the lamellar interface. A weakened dermo-epidermal junction is a possible intermediate stage in the pathophysiology of bovine sole ulceration at the typical site.

  13. A MEMS lamination technology based on sequential multilayer electrodeposition

    SciTech Connect

    Kim, M; Kim, J; Herrault, F; Schafer, R; Allen, MG

    2013-08-06

    A MEMS lamination technology based on sequential multilayer electrodeposition is presented. The process comprises three main steps: (1) automated sequential electrodeposition of permalloy (Ni80Fe20) structural and copper sacrificial layers to form multilayer structures of significant total thickness; (2) fabrication of polymeric anchor structures through the thickness of the multilayer structures and (3) selective removal of copper. The resulting structure is a set of air-insulated permalloy laminations, the separation of which is sustained by insulating polymeric anchor structures. Individual laminations have precisely controllable thicknesses ranging from 500 nm to 5 mu m, and each lamination layer is electrically isolated from adjacent layers by narrow air gaps of similar scale. In addition to air, interlamination insulators based on polymers are investigated. Interlamination air gaps with very high aspect ratio (>1:100) can be filled with polyvinylalcohol and polydimethylsiloxane. The laminated structures are characterized using scanning electron microscopy and atomic force microscopy to directly examine properties such as the roughness and the thickness uniformity of the layers. In addition, the quality of the electrical insulation between the laminations is evaluated by quantifying the eddy current within the sample as a function of frequency. Fabricated laminations are comprised of uniform, smooth (surface roughness < 100 nm) layers with effective electrical insulation for all layer thicknesses and insulator approaches studied. Such highly laminated structures have potential uses ranging from energy conversion to applications where composite materials with highly anisotropic mechanical or thermal properties are required.

  14. Self-heating forecasting for thick laminate specimens in fatigue

    NASA Astrophysics Data System (ADS)

    Lahuerta, F.; Westphal, T.; Nijssen, R. P. L.

    2014-12-01

    Thick laminate sections can be found from the tip to the root in most common wind turbine blade designs. Obtaining accurate and reliable design data for thick laminates is subject of investigations, which include experiments on thick laminate coupons. Due to the poor thermal conductivity properties of composites and the material self-heating that occurs during the fatigue loading, high temperature gradients may appear through the laminate thickness. In the case of thick laminates in high load regimes, the core temperature might influence the mechanical properties, leading to premature failures. In the present work a method to forecast the self-heating of thick laminates in fatigue loading is presented. The mechanical loading is related with the laminate self-heating, via the cyclic strain energy and the energy loss ratio. Based on this internal volumetric heat load a thermal model is built and solved to obtain the temperature distribution in the transient state. Based on experimental measurements of the energy loss factor for 10mm thick coupons, the method is described and the resulting predictions are compared with experimental surface temperature measurements on 10 and 30mm UD thick laminate specimens.

  15. Laminated lumber may be more profitable than sawn lumber

    Treesearch

    P. Koch

    1976-01-01

    By laminating 1/4-in. rotary-cut veneer into structural lumber, manufacturers can expand lumber output by at least 30% without increasing volume logged. The idea merits intensive study. Manufacturing plus raw material costs should total about $142/Mbf; sales price for desirable widths and lengths of the strong laminated product should approach or exceed $200/Mbf.

  16. A Laminated Track for the Inductrack System: Theory and Experiment

    SciTech Connect

    Post, R F; Hoburg, J F

    2004-01-12

    A laminated structure, composed of stacks of thin conducting sheets, has several advantages over a litz-wire ladder as the ''track'' wherein levitating currents are induced by a permanent magnet array on a moving vehicle. Modeling and experimental results for the laminated track are described and evaluated in this paper.

  17. Residual stresses and their effects in composite laminates

    NASA Technical Reports Server (NTRS)

    Hahn, H. T.; Hwang, D. G.

    1983-01-01

    Residual stresses in composite laminates are caused by the anisotropy in expansional properties of constituent unidirectional plies. The effect of these residual stresses on dimensional stability is studied through the warping of unsymmetric (0 sub 4/90 sub 4)sub T graphite/epoxy laminates while their effect on ply failure is analyzed for (0/90)sub 2s Kevlar 49/epoxy laminate. The classical laminated plate theory is used to predict the warping of small and large panels. The change of warping does not indicate a noticeable stress relaxation at 75 C while it is very sensitive to moisture content and hence to environment. A prolonged gellation at the initial cure temperature reduces residual stresses while postcure does not. The matrix/interface cracking in dry (0/90)sub 2s Kevlar 49/epoxy laminate is shown to be the result of the residual stress exceeding the transverse strength.

  18. Deposition of laminated shale: A field and experimental study

    NASA Astrophysics Data System (ADS)

    Carey, Daniel L.; Roy, David C.

    1985-03-01

    Intermittently laminated shale of the Jemtland Formation in Maine is characterized by thin lenticular silt segregations interlaced with argillaceous and organic material (including graptolites). This shale is thinly interbedded with nonlaminated shale, siltstone, and thicker turbidite graywacke beds. Experiments suggest that the intermittently laminated shale was deposited by silt/clay-laden currents and may have been part of an upward turbidite progression from parallel-laminated silt (>60% silt), through intermittently laminated mud (40 to 60% silt), to nonlaminated mud (20 to 40% silt). Intermittently laminated mud may be produced from silt/clay flows that are: decelerating at a constant silt content, losing silt at constant velocity; or both decelerating and losing silt.

  19. The strength of laminated composite materials under repeated impact loading

    NASA Technical Reports Server (NTRS)

    Rotem, Assa

    1988-01-01

    When low velocity and energy impact is exerted on a laminated composite material, in a perpendicular direction to the plane of the laminate, invisible damage may develop. It is shown analytically and experimentally that the invisible damage occurs during the first stage of contact between the impactor and the laminate and is a result of the contact stresses. However, the residual flexural strength changes only slightly, because it depends mainly on the outer layers, and these remain undamaged. Repeated impact intensifies the damage inside the laminate and causes larger bending under equivalent impact load. Finally, when the damage is most severe, even though it is still invisible, the laminate fails because of bending on the tension side. If the repeated impact is halted before final fracture occurs the residual strength and modulus would decrease by a certain amount.

  20. The mechanical behavior of GLARE laminates for aircraft structures

    NASA Astrophysics Data System (ADS)

    Wu, Guocai; Yang, J.-M.

    2005-01-01

    GLARE (glass-reinforced aluminum laminate) is a new class of fiber metal laminates for advanced aerospace structural applications. It consists of thin aluminum sheets bonded together with unidirectional or biaxially reinforced adhesive prepreg of high-strength glass fibers. GLARE laminates offer a unique combination of properties such as outstanding fatigue resistance, high specific static properties, excellent impact resistance, good residual and blunt notch strength, flame resistance and corrosion properties, and ease of manufacture and repair. GLARE laminates can be tailored to suit a wide variety of applications by varying the fiber/resin system, the alloy type and thickness, stacking sequence, fiber orientation, surface pretreatment technique, etc. This article presents a comprehensive overview of the mechanical properties of various GLARE laminates under different loading conditions.

  1. Matrix cracking in laminated composites under monotonic and cyclic loadings

    NASA Technical Reports Server (NTRS)

    Allen, David H.; Lee, Jong-Won

    1991-01-01

    An analytical model based on the internal state variable (ISV) concept and the strain energy method is proposed for characterizing the monotonic and cyclic response of laminated composites containing matrix cracks. A modified constitution is formulated for angle-ply laminates under general in-plane mechanical loading and constant temperature change. A monotonic matrix cracking criterion is developed for predicting the crack density in cross-ply laminates as a function of the applied laminate axial stress. An initial formulation for a cyclic matrix cracking criterion for cross-ply laminates is also discussed. For the monotonic loading case, a number of experimental data and well-known models are compared with the present study for validating the practical applicability of the ISV approach.

  2. Design of composite laminates by a Monte Carlo method

    NASA Astrophysics Data System (ADS)

    Fang, Chin; Springer, George S.

    1993-01-01

    A Monte Carlo procedure was developed for optimizing symmetric fiber reinforced composite laminates such that the weight is minimum and the Tsai-Wu strength failure criterion is satisfied in each ply. The laminate may consist of several materials including an idealized core, and may be subjected to several sets of combined in-plane and bending loads. The procedure yields the number of plies, the fiber orientation, and the material of each ply and the material and thickness of the core. A user friendly computer code was written for performing the numerical calculations. Laminates optimized by the code were compared to laminates resulting from existing optimization methods. These comparisons showed that the present Monte Carlo procedure is a useful and efficient tool for the design of composite laminates.

  3. The strength of laminated composite materials under repeated impact loading

    NASA Technical Reports Server (NTRS)

    Rotem, Assa

    1988-01-01

    When low velocity and energy impact is exerted on a laminated composite material, in a perpendicular direction to the plane of the laminate, invisible damage may develop. It is shown analytically and experimentally that the invisible damage occurs during the first stage of contact between the impactor and the laminate and is a result of the contact stresses. However, the residual flexural strength changes only slightly, because it depends mainly on the outer layers, and these remain undamaged. Repeated impact intensifies the damage inside the laminate and causes larger bending under equivalent impact load. Finally, when the damage is most severe, even though it is still invisible, the laminate fails because of bending on the tension side. If the repeated impact is halted before final fracture occurs the residual strength and modulus would decrease by a certain amount.

  4. Residual stresses and their effects in composite laminates

    NASA Technical Reports Server (NTRS)

    Hahn, H. T.; Hwang, D. G.

    1983-01-01

    Residual stresses in composite laminates are caused by the anisotropy in expansional properties of constituent unidirectional plies. The effect of these residual stresses on dimensional stability is studied through the warping of unsymmetric (0 sub 4/90 sub 4)sub T graphite/epoxy laminates while their effect on ply failure is analyzed for (0/90)sub 2s Kevlar 49/epoxy laminate. The classical laminated plate theory is used to predict the warping of small and large panels. The change of warping does not indicate a noticeable stress relaxation at 75 C while it is very sensitive to moisture content and hence to environment. A prolonged gellation at the initial cure temperature reduces residual stresses while postcure does not. The matrix/interface cracking in dry (0/90)sub 2s Kevlar 49/epoxy laminate is shown to be the result of the residual stress exceeding the transverse strength.

  5. Vibration suppression of composite laminated plate with nonlinear energy sink

    NASA Astrophysics Data System (ADS)

    Zhang, Ye-Wei; Zhang, Hao; Hou, Shuai; Xu, Ke-Fan; Chen, Li-Qun

    2016-06-01

    The composite laminated plate is widely used in supersonic aircraft. So, there are many researches about the vibration suppression of composite laminated plate. In this paper, nonlinear energy sink (NES) as an effective method to suppress vibration is studied. The coupled partial differential governing equations of the composite laminated plate with the nonlinear energy sink (NES) are established by using the Hamilton principle. The fourth-order Galerkin discrete method is used to truncate the partial differential equations, which are solved by numerical integration method. Meanwhile study about the precise effectiveness of the nonlinear energy sink (NES) by discussing the different installation location of the nonlinear energy sink (NES) at the same speed. The results indicate that the nonlinear energy sink (NES) can significantly suppress the severe vibration of the composite laminated plate with speed wind loadings in to protect the composite laminated plate from excessive vibration.

  6. Matrix cracking in laminated composites under monotonic and cyclic loadings

    NASA Technical Reports Server (NTRS)

    Allen, David H.; Lee, Jong-Won

    1991-01-01

    An analytical model based on the internal state variable (ISV) concept and the strain energy method is proposed for characterizing the monotonic and cyclic response of laminated composites containing matrix cracks. A modified constitution is formulated for angle-ply laminates under general in-plane mechanical loading and constant temperature change. A monotonic matrix cracking criterion is developed for predicting the crack density in cross-ply laminates as a function of the applied laminate axial stress. An initial formulation for a cyclic matrix cracking criterion for cross-ply laminates is also discussed. For the monotonic loading case, a number of experimental data and well-known models are compared with the present study for validating the practical applicability of the ISV approach.

  7. Geometrically nonlinear behavior of piezoelectric laminated plates

    NASA Astrophysics Data System (ADS)

    Rabinovitch, Oded

    2005-08-01

    The geometrically nonlinear behavior of piezo-laminated plates actuated with isotropic or anisotropic piezoelectric layers is analytically investigated. The analytical model is derived using the variational principle of virtual work along with the lamination and plate theories, the von Karman large displacement and moderate rotation kinematic relations, and the anisotropic piezoelectric constitutive laws. A solution strategy that combines the approach of the method of lines, the advantages of the finite element concept, and the variational formulation is developed. This approach yields a set of nonlinear ordinary differential equations with nonlinear boundary conditions, which are solved using the multiple-shooting method. Convergence and verification of the model are examined through comparison with linear and nonlinear results of other approximation methods. The nonlinear response of two active plate structures is investigated numerically. The first plate is actuated in bending using monolithic piezoceramic layers and the second one is actuated in twist using macro-fiber composites. The results quantitatively reveal the complicated in-plane stress state associated with the piezoelectric actuation and the geometrically nonlinear coupling of the in-plane and out-of-plane responses of the plate. The influence of the nonlinear effects ranges from significant stiffening in certain combinations of electrical loads and boundary conditions to amplifications of the induced deflections in others. The paper closes with a summary and conclusions.

  8. Free Vibration of Uncertain Unsymmetrically Laminated Beams

    NASA Technical Reports Server (NTRS)

    Kapania, Rakesh K.; Goyal, Vijay K.

    2001-01-01

    Monte Carlo Simulation and Stochastic FEA are used to predict randomness in the free vibration response of thin unsymmetrically laminated beams. For the present study, it is assumed that randomness in the response is only caused by uncertainties in the ply orientations. The ply orientations may become random or uncertain during the manufacturing process. A new 16-dof beam element, based on the first-order shear deformation beam theory, is used to study the stochastic nature of the natural frequencies. Using variational principles, the element stiffness matrix and mass matrix are obtained through analytical integration. Using a random sequence a large data set is generated, containing possible random ply-orientations. This data is assumed to be symmetric. The stochastic-based finite element model for free vibrations predicts the relation between the randomness in fundamental natural frequencies and the randomness in ply-orientation. The sensitivity derivatives are calculated numerically through an exact formulation. The squared fundamental natural frequencies are expressed in terms of deterministic and probabilistic quantities, allowing to determine how sensitive they are to variations in ply angles. The predicted mean-valued fundamental natural frequency squared and the variance of the present model are in good agreement with Monte Carlo Simulation. Results, also, show that variations between plus or minus 5 degrees in ply-angles can affect free vibration response of unsymmetrically and symmetrically laminated beams.

  9. Residual stress characterization for laminated composites

    NASA Astrophysics Data System (ADS)

    Liu, Shao-Chun

    With increasing applications of advanced laminated composites, process-induced residual stress has drawn more and more attention in recent years. Efforts have been devoted to understanding residual stress both quantitatively and qualitatively. In the current study, a novel technique called the Cure Referencing Method was developed which has the capability for measuring the residual stress on the symmetric laminated composite plates. It can also differentiate residual stress into two components: one is due to the mismatch of the coefficient of thermal expansion, the other is caused by the matrix chemical curing shrinkage. The chemical curing shrinkage of the polymer matrix was investigated in further detail. A technique was developed to measure the post-gel chemical curing shrinkage which is the portion of curing shrinkage that really induces the residual stress in the polymer matrix composites. Time-dependent material property is another issue associated with polymer matrix composite materials. The data of several short-term tensile creep tests run at different temperature were used to construct a linear viscoelastic: model for describing the behavior of the composites over a long period of time. It was found that physical aging of the polymer matrix needs to be taken into account in order to have a more accurate representation of the long-term behavior. A fair agreement was obtained between the result of the long-term creep test and the master curve constructed from several momentary creep tests.

  10. Interlaminar interaction in paper thermoplastic laminate composites

    NASA Astrophysics Data System (ADS)

    Prambauer, M.; Paulik, C.; Burgstaller, C.

    2016-07-01

    Bio-based composites are a research topic since several decades, which aims for sustainable and durable materials. In the scope of this research, many different sources for biobased reinforcements have been investigated. Typical issues associated with the use of such are property variations due to cultivation area and climate, besides the influences of the type, pretreatment and fibre geometry. Another issue can be the availability of such natural fibres. Due to these reasons, we started using paper sheets as reinforcements in laminate composites with thermoplastic materials. In preliminary studies with polypropylene composites, we found good mechanical properties, even higher than could be expected by estimating the composite properties from the constituents by applying simple rule of mixtures type models. We suspect, besides some effect of paper compaction, interlaminar effects to be the reason for this. Therefore, the aim of this work is to investigate the effects of the interfacial interaction on the different paper laminate properties due to different matrix polymers. For this work, we used polypropylene, polyamide 6 and 12 as well as polystyrene. Composites were produced via compression moulding and samples for mechanical testing and density evaluation were cut from the moulded plates. The results from mechanical tests show, that there is a reinforcing effect, regardless of matrix polymer used. Simple rule of mixtures evaluations show, that the different matrices exhibit different degrees of interaction, based on their chemical structure. In addition, also influences due to processing were found.

  11. Development of a heterogeneous laminating resin system

    NASA Technical Reports Server (NTRS)

    Biermann, T. F.; Hopper, L. C.

    1985-01-01

    The factors which effect the impact resistance of laminating resin systems and yet retain equivalent performance with the conventional 450 K curing epoxy matrix systems in other areas were studied. Formulation work was conducted on two systems, an all-epoxy and an epoxy/bismaleimide, to gain fundamental information on the effect formulation changes have upon neat resin and composite properties. The all-epoxy work involved formulations with various amounts and combinations of eight different epoxy resins, four different hardeners, fifteen different toughening agents, a filler, and a catalyst. The epoxy/bismaleimide effort improved formulations with various amounts and combinations of nine different resins, four different hardeners, eight different toughening agents, four different catalysts, and a filler. When a formulation appeared to offer the proper combination of properties required for a laminating resin Celion 3K-70P fabric was prepregged. Initial screening tests on composites primarily involved Gardner type impact and measurement of short beam shear strengths under dry and hot/wet conditions.

  12. Nuclear envelope structural proteins facilitate nuclear shape changes accompanying embryonic differentiation and fidelity of gene expression.

    PubMed

    Smith, Elizabeth R; Meng, Yue; Moore, Robert; Tse, Jeffrey D; Xu, Arn G; Xu, Xiang-Xi

    2017-01-14

    Nuclear size and shape are specific to a cell type, function, and location, and can serve as indicators of disease and development. We previously found that lamin A/C and associated nuclear envelope structural proteins were upregulated when murine embryonic stem (ES) cells differentiated to primitive endoderm cells. Here we further investigated the morphological changes of nuclei that accompany this differentiation. The nuclei of undifferentiated wild type cells were found shaped as flattened, irregular ovals, whereas nuclei of Gata4-positive endoderm cells were more spherical, less flattened, and with a slightly reduced volume. The morphological change was confirmed in the trophectoderm and primitive endoderm lineages of E4.5 blastocysts, compared to larger and more irregularly shaped of the nuclei of the inner cell mass. We established ES cells genetically null for the nuclear lamina proteins lamin A/C or the inner nuclear envelope protein emerin, or compound mutant for both lamin A/C and emerin. ES cells deficient in lamin A/C differentiated to endoderm but less efficiently, and the nuclei remained flattened and failed to condense. The size and shape of emerin-deficient nuclei also remained uncondensed after treatment with RA. The emerin/lamin A/C double knockout ES cells failed to differentiate to endoderm cells, though the nuclei condensed but retained a generally flattened ellipsoid shape. Additionally, ES cells deficient for lamin A/C and/or emerin had compromised ability to undergo endoderm differentiation, where the differentiating cells often exhibited coexpression of pluripotent and differentiation markers, such as Oct3/4 and Gata4, respectively, indicating an infidelity of gene regulation. The results suggest that changes in nuclear size and shape, which are mediated by nuclear envelope structural proteins lamin A/C and/or emerin, also impact gene regulation and lineage differentiation in early embryos. Nevertheless, mice lacking both lamin A/C and

  13. Failure and deformation analyses of smart laminated composites

    NASA Astrophysics Data System (ADS)

    Hasan, Z.; Muliana, A.

    2012-09-01

    The present study focuses on the failure analysis and shape control of smart composite laminates under coupled thermal (hygro), electric, and mechanical stimuli. A linear thermo(hygro)electroelastic constitutive model for transversely isotropic materials is used for each ply in the composite laminate and for the piezoelectric materials that are integrated with laminates of the composite. Piezoelectric materials, such as lead zirconate titanate, and piezoelectric fiber composites, such as an active fiber composite or a microfiber composite, are considered as actuators for controlling unwanted bending deformations to avoid failure in such composite laminates. Due to the high stress concentrations at the interfaces between an active layer and the host structure, which may cause debonding, embedded actuators in which the active material is placed as part of the plies to form geometrically continuous plies are considered in order to minimize the stress concentration while improving the actuation capability. The first-ply failure and the ultimate laminate failure criteria of composite laminates are used to predict the failure stress and mode of the smart composite laminates, where commonly known macroscopic failure criteria, such as the Tsai-Hill, Tsai-Wu, and maximum stress criteria, are employed for each lamina. Piezoelectric materials can be used to prevent the failure from hygrothermal and mechanical loadings by applying an electric voltage in order to counteract laminate deformations. Based on the deformation and failure analyzes of smart composite laminates having various stacking sequences, fiber and matrix constituents, and piezoelectric materials, we could estimate the overall properties and failure envelopes of the laminates, which is useful in the preliminary design of smart composite structures.

  14. Buckling analysis of laminated thin shells in a hot environment

    NASA Technical Reports Server (NTRS)

    Gotsis, Pascal K.; Guptil, James D.

    1993-01-01

    Results are presented of parametric studies to assess the effects of various parameters on the buckling behavior of angle-ply, laminated thin shells in a hot environment. These results were obtained by using a three-dimensional finite element analysis. An angle-ply, laminated thin shell with fiber orientation of (theta/-theta)(sub 2) was subjected to compressive mechanical loads. The laminated thin shell had a cylindrical geometry. The laminate contained T300 graphite fibers embedded in an intermediate-modulus, high-strength (IMHS) matrix. The fiber volume fraction was 55 percent and the moisture content was 2 percent. The residual stresses induced into the laminate structure during the curing were taken into account. Parametric studies were performed to examine the effect on the critical buckling load of the following parameters: cylinder length and thickness, internal hydrostatic pressure, different ply thicknesses, different temperature profiles through the thickness of the structure, and different lay up configurations and fiber volume fractions. In conjunction with these parameters the ply orientation was varied from 0 deg to 90 deg. Seven ply angles were examined: 0 deg, 15 deg, 30 deg, 45 deg, 60 deg, 75 deg, and 90 deg. The results show that the ply angle theta and the laminate thickness had significant effects on the critical buckling load. The fiber volume fraction, the fiber orientations, and the internal hydrostatic pressure had important effects on the critical buckling load. The cylinder length had a moderate influence on the buckling load. The thin shell with (theta/-theta)(sub 2) or (theta/-theta)(sub s) angle-ply laminate had better buckling-load performance than the thin shell with (theta)(sub 4) angle-ply laminate. The temperature profiles through the laminate thickness and various laminates with the different ply thicknesses has insignificant effects on the buckling behavior of the thin shells.

  15. Interlaminate Deformation in Thermoplastic Composite Laminates: Experimental-Numerical Correlation

    NASA Astrophysics Data System (ADS)

    Shen, M.; Tong, J.; Wang, S.; Fang, Y.

    2010-06-01

    The interlaminar deformation behaviors of thermoplastic AS4/PEEK composite laminates subjected to static tensile loading are investigated by means of microscopic moiré interferometry with high spatial resolution. The fully threedimensional orthotropic elastic-plastic analysis of interlaminar deformation for the thermoplastic laminates is developed in this paper, and used to simulate the stress-strain curves of tensile experiment for its angle-ply laminates. Under uniaxial tensile loading, the 3D orthotropic elastic-plastic FE analysis and microscopic moiré interferometry of interlaminar deformations are carried out for the [±25]S4 laminates. The quantitative local-filed experimental results of interlaminar shear strain and displacements at freeedge surface of the laminate are compared with corresponding numerical results of the orthotropic elastic-plastic FE model. It is indicated that the numerical tensile stressstrain curves of angle-ply laminates computed with 3D orthotropic elastic-plastic model are agree with experimental results. The numerical interlaminar displacement U and shear strain γxz are also consistent with the experimental results obtained by moiré interferometry. It is expected the elastic-plastic interlaminar stresses and deformations analysis for the optimal design and application of AS4/PEEK laminates and its structures.

  16. Low cost and high performance screen laminate regenerator matrix

    NASA Astrophysics Data System (ADS)

    Bin-Nun, Uri; Manitakos, Dan

    2004-06-01

    A laminate screen matrix regenerator with 47 elements has been designed, analyzed, fabricated and tested. The laminate was fabricated from stainless steel screen sheets that were stacked on top of each other at certain angular orientation and then bonded at high temperature and pressure environment utilizing a sintering process. This laminate is a porous structure media with highly repeatable properties that can be controlled by varying mesh size, weave type, wire size and laminate sheet to sheet orientation. The flow direction in relation to the weave plan can be varied by cutting a cylindrical or rectangular laminate element along or across the weave. The regenerator flow resistance, thermal conductance losses, dead volume, surface area and heat transfer coefficient are analyzed. Regenerator cost and performance comparison data between the conventional widely used method of stacked screens and the new stacked laminate matrix regenerator is discussed. Also, a square stainless steel screen laminate was manufactured in a way which permits gas to flow along the screen wire instead of across it.

  17. Bending strength of shallow glued-laminated beams of a uniform grade

    Treesearch

    Catherine M. Marx; Russell C. Moody

    1981-01-01

    Ninety glued-laminated Douglas-fir or southern pine beams of a uniform grade with 2-, 4-, or 6-laminations were evaluated in static bending tests. No specially graded tension laminations or end joints were used. The purpose of the tests was to determine which of three present design criteria best predict near minimum bending strength values for shallow glued-laminated...

  18. LamLum : a tool for evaluating the financial feasibility of laminated lumber plants

    Treesearch

    E.M. (Ted) Bilek; John F. Hunt

    2006-01-01

    A spreadsheet-based computer program called LamLum was created to analyze the economics of value- added laminated lumber manufacturing facilities. Such facilities manufacture laminations, typically from lower grades of structural lumber, then glue these laminations together to make various types of higher value laminated lumber products. This report provides the...

  19. Scintigraphic evaluation of digital circulation during the developmental and acute phases of equine laminitis

    SciTech Connect

    Trout, D.R.

    1987-01-01

    Using nuclear isotopic imaging, digital circulation was sequentially evaluated at 24-hour intervals in 11 control horses and in 9 horses affected with acute laminitis, created by administration of a high-starch ration. Following intra-arterial injection of /sup 99m/Tc macroaggregated albumin into the brachiocephalic trunk, a gamma camera and dedicated nuclear medicine computer were used to acquire static images of the right front foot. Dynamic vascular-phase and static interstitial-phase images were also obtained after jugular vein injection of /sup 99m/Tc diethylenetriamine pentaacetic acid. These procedures were performed on standing horses, using either minimal or no tranquilization. The images were quantitatively analyzed for parameters indicative of circulation to the foot as a whole and to specific regions of interest within the foot. There was no evidence of reduced total blood flow to the lamellae during either the developmental or acute phases of laminitis. Although total flow tended to increase throughout the peripheral/external regions of the foot, statistically significant elevations were consistently present only within the lamellae. Changes indicative of decreased total blood flow were noted in the central/internal regions of the foot. These alterations usually occurred coincident with or after the onset of clinical lameness.

  20. β-catenin is essential for lamination but not neurogenesis in mouse retinal development

    PubMed Central

    Fu, Xueyao; Sun, Hongxia; Klein, William H.; Mu, Xiuqian

    2007-01-01

    During vertebrate retinal development, the seven retinal cell types differentiate sequentially from a single population of retinal progenitor cells (RPCs) and organize themselves into a distinct laminar structure. The purpose of this study was to determine whether β-catenin, which functions both as a nuclear effector for the canonical Wnt signaling pathway and as a regulator of cell adhesion, is required for retinal neurogenesis or lamination. We used the Cre-loxP system to either eliminate β-catenin or to express a constitutively active form during retinal neurogenesis. Eliminating β-catenin did not affect cell differentiation, but did result in the loss of the radial arrangement of RPCs and caused abnormal migration of differentiated neurons. As a result, the laminar structure was massively disrupted in β-catenin-null retinas, although all retinal cell types still formed. In contrast to other neural tissues, eliminating β-catenin did not significantly reduce the proliferation rate of RPCs; likewise, activating β-catenin ectopically in RPCs did not result in overproliferation, but loss of neural retinal identity. These results indicate that β-catenin is essential during retinal neurogenesis as a regulator of cell adhesion but not as a nuclear effector of the canonical Wnt signaling pathway. The results further imply that retinal lamination and retinal cell differentiation are genetically separable processes. PMID:16959241

  1. Formaldehyde and TVOC emission behavior of laminate flooring by structure of laminate flooring and heating condition.

    PubMed

    An, Jae-Yoon; Kim, Sumin; Kim, Hyun-Joong

    2011-03-15

    Formaldehyde was measured with a desiccator, a 20 L chamber and the FLEC method. The formaldehyde emission rate from laminate was the highest at 32 °C using the desiccator, which then decreased with time. The formaldehyde emission using the 20 L small chamber and FLEC showed a similar tendency. There was a strong correlation between the formaldehyde and total volatile organic compounds (TVOCs) with both types of floorings using the two different methods. The formaldehyde emission rate and TVOC results were higher when tested using the FLEC method than with the 20 L small chamber method. The emission rate was affected by the joint edge length in laminate flooring. Toluene, ethylbenzene and xylene were the main VOCs emitted from laminate flooring, and there were more unidentified VOCs emitted than identified VOCs. The samples heated with a floor heating system emitted more formaldehyde than those heated using an air circulation system due to the temperature difference between the bottom panel and flooring. The TVOC emission level of the samples was higher when an air circulation system was used than when a floor heating system was used due to the high ventilation rate.

  2. Nuclear envelopathies: a complex LINC between nuclear envelope and pathology.

    PubMed

    Janin, Alexandre; Bauer, Delphine; Ratti, Francesca; Millat, Gilles; Méjat, Alexandre

    2017-08-30

    Since the identification of the first disease causing mutation in the gene coding for emerin, a transmembrane protein of the inner nuclear membrane, hundreds of mutations and variants have been found in genes encoding for nuclear envelope components. These proteins can be part of the inner nuclear membrane (INM), such as emerin or SUN proteins, outer nuclear membrane (ONM), such as Nesprins, or the nuclear lamina, such as lamins A and C. However, they physically interact with each other to insure the nuclear envelope integrity and mediate the interactions of the nuclear envelope with both the genome, on the inner side, and the cytoskeleton, on the outer side. The core of this complex, called LINC (LInker of Nucleoskeleton to Cytoskeleton) is composed of KASH and SUN homology domain proteins. SUN proteins are INM proteins which interact with lamins by their N-terminal domain and with the KASH domain of nesprins located in the ONM by their C-terminal domain.Although most of these proteins are ubiquitously expressed, their mutations have been associated with a large number of clinically unrelated pathologies affecting specific tissues. Moreover, variants in SUN proteins have been found to modulate the severity of diseases induced by mutations in other LINC components or interactors. For these reasons, the diagnosis and the identification of the molecular explanation of "nuclear envelopathies" is currently challenging.The aim of this review is to summarize the human diseases caused by mutations in genes coding for INM proteins, nuclear lamina, and ONM proteins, and to discuss their potential physiopathological mechanisms that could explain the large spectrum of observed symptoms.

  3. Nuclear condensation during mouse erythropoiesis requires caspase-3-mediated nuclear opening

    PubMed Central

    Zhao, Baobing; Mei, Yang; Schipma, Matthew J; Roth, Eric Wayne; Bleher, Reiner; Rappoport, Joshua Z.; Wickrema, Amittha; Yang, Jing; Ji, Peng

    2016-01-01

    SUMMARY Mammalian erythropoiesis involves chromatin condensation that is initiated in the early stage of terminal differentiation. The mechanisms of chromatin condensation during erythropoiesis are unclear. Here, we show that the mouse erythroblast forms large, transient, and recurrent nuclear openings that coincide with the condensation process. The opening lacks nuclear lamina, nuclear pore complexes, and nuclear membrane, but it is distinct from nuclear envelope changes that occur during apoptosis and mitosis. A fraction of the major histones are released from the nuclear opening and degraded in the cytoplasm. We demonstrate that caspase-3 is required for the nuclear opening formation throughout terminal erythropoiesis. Loss of caspase-3 or ectopic expression of a caspase-3 non-cleavable lamin B mutant blocks nuclear opening formation, histone release, chromatin condensation, and terminal erythroid differentiation. We conclude that caspase-3-mediated nuclear opening formation accompanied by histone release from the opening is a critical step towards chromatin condensation during erythropoiesis in mice. PMID:26954545

  4. Chronic laminitis: strategic hoof wall resection.

    PubMed

    Rucker, Amy

    2010-04-01

    In the chronic-laminitic foot, severe soft-tissue compression and compromised circulation can result in osteitis and sepsis at the margin of the distal phalanx. Resultant inflammation and sepsis may cause the coronary corium to swell, drain, or separate from the hoof capsule, usually within 8 weeks of laminitis onset. Slow-onset cases of soft-tissue impingement can develop secondary to distal phalanx displacement due to lack of wall attachment. With either presentation, partial upper wall resection is required to reverse compression and vascular impingement by the hoof capsule. If the pathology is not overwhelming, the area reepithelializes and grows attached tubular horn. Firm bandaging and restricted exercise until tubular horn has regrown enhances recovery and the return of a strong hoof.

  5. The digital pathologies of chronic laminitis.

    PubMed

    Grosenbaugh, D A; Morgan, S J; Hood, D M

    1999-08-01

    This review indicates that the patient-to-patient uniqueness commonly seen in chronic laminitis represents the variable presence of the digital pathologies. Although some degree of mechanical failure is always present, the secondary metabolic and growth dysplasias, vascular pathologies, and sepsis may or may not be evident. The presence and severity of these pathologies appear to have a more significant impact on the prognosis of individual cases than does the displacement of the distal phalanx. It should be reiterated that it is often the combined presence of these individual pathologies that gives rise to the patient that is totally refractory to treatment. In the absence of these pathologies, many horses with significant displacement of the distal phalanx are not in pain and are not in need of treatment. It thus follows that a key to the improved rehabilitation of difficult patients is focusing research on the physiopathology and diagnosis of these nonmechanical problems.

  6. Statistical fatigue of unnotched composite laminates

    NASA Technical Reports Server (NTRS)

    Yang, J. N.; Jones, D. L.

    1980-01-01

    A general analytical, phenomenological model is developed for characterizing fatigue damage accumulation in unnotched composite laminates, and experimental results are used to evaluate the model. The fatigue model, which is developed for various types of cyclic loadings, is based on the assumption that the residual strength reduction is a monotonically decreasing function of the fatigue life. Parameters are determined from baseline test data consisting of one set of residual strength data, one set of static strength data, and one set of constant amplitude fatigue life data. Once the parameters are determined, the model is able to predict: (1) the statistical distribution of the fatigue life, and the residual strength under constant and variable amplitude loads, and (2) the effect of periodic proof tests, without any additional experimental data. Experimental test results show that predictions made from the present model agree quite well with the experimental results. The model is also applicable to fatigue of bonded or bolted composite joints.

  7. Compression failure of angle-ply laminates

    NASA Technical Reports Server (NTRS)

    Peel, L. D.; Hyer, M. W.; Shuart, M. J.

    1992-01-01

    Test results from the compression loading of (+ or - Theta/ - or + Theta)(sub 6s) angle-ply IM7-8551-7a specimens, 0 less than or = Theta less than or = 90 degs, are presented. The observed failure strengths and modes are discussed, and typical stress-strain relations shown. Using classical lamination theory and the maximum stress criterion, an attempt is made to predict failure stress as a function of Theta. This attempt results in poor correlation with test results and thus a more advanced model is used. The model, which is based on a geometrically nonlinear theory, and which was taken from previous work, includes the influence of observed layer waviness. The waviness is described by the wave length and the wave amplitude. The theory is briefly described and results from the theory are correlated with test results. It is shown that by using levels of waviness observed in the specimens, the correlation between predictions and observations is good.

  8. Permeability After Impact Testing of Composite Laminates

    NASA Technical Reports Server (NTRS)

    Nettles, A.T.; Munafo, Paul (Technical Monitor)

    2002-01-01

    Since composite laminates are beginning to be identified for use in reusable launch vehicle propulsion systems, an understanding of their permeance is needed. A foreign object impact event can cause a localized area of permeability (leakage) in a polymer matrix composite and it is the aim of this study to assess a method of quantifying permeability-after-impact results. A simple test apparatus is presented and variables that could affect the measured values of permeability-after-impact were assessed. Once it was determined that valid numbers were being measured, a fiber/resin system was impacted at various impact levels and the resulting permeability measured, first with a leak check solution (qualitative) then using the new apparatus (quantitative). The results showed that as the impact level increased, so did the measured leakage. As the pressure to the specimen was increased, the leak rate was seen to increase in a non-linear fashion for almost all of the specimens tested.

  9. Permeability After Impact Testing of Composite Laminates

    NASA Technical Reports Server (NTRS)

    Nettles, Alan T.

    2003-01-01

    Since composite laminates are beginning to be identified for use in reusable launch vehicle propulsion systems, an understanding of their permeance is needed. A foreign object impact event can cause a localized area of permeability (leakage) in a polymer matrix composite and it is the aim of this study to assess a method of quantifying permeability-after-impact results. A simple test apparatus is presented and variables that could affect the measured values of permeability-after-impact were assessed. Once it was determined that valid numbers were being measured, a fiber/resin system was impacted at various impact levels and the resulting permeability measured, first with a leak check solution (qualitative) then using the new apparatus (quantitative). The results showed that as the impact level increased, so did the measured leakage. As the pressure to the specimen was increased, the leak rate was seen to increase in a non-linear fashion for almost all of the specimens tested.

  10. Reliability analysis of ceramic matrix composite laminates

    NASA Technical Reports Server (NTRS)

    Thomas, David J.; Wetherhold, Robert C.

    1991-01-01

    At a macroscopic level, a composite lamina may be considered as a homogeneous orthotropic solid whose directional strengths are random variables. Incorporation of these random variable strengths into failure models, either interactive or non-interactive, allows for the evaluation of the lamina reliability under a given stress state. Using a non-interactive criterion for demonstration purposes, laminate reliabilities are calculated assuming previously established load sharing rules for the redistribution of load as the failure of laminae occur. The matrix cracking predicted by ACK theory is modeled to allow a loss of stiffness in the fiber direction. The subsequent failure in the fiber direction is controlled by a modified bundle theory. Results using this modified bundle model are compared with previous models which did not permit separate consideration of matrix cracking, as well as to results obtained from experimental data.

  11. The Creep of Laminated Synthetic Resin Plastics

    NASA Technical Reports Server (NTRS)

    Perkuhn, H

    1941-01-01

    The long-time loading strength of a number of laminated synthetic resin plastics was ascertained and the effect of molding pressure and resin content determined. The best value was observed with a 30 to 40 percent resin content. The long-time loading strength also increases with increasing molding pressure up to 250 kg/cm(exp 2); a further rise in pressure affords no further substantial improvement. The creep strength is defined as the load which in the hundredth hour of loading produces a rate of elongation of 5 X 10(exp -4) percent per hour. The creep strength values of different materials were determined and tabulated. The effect of humidity during long-term tests is pointed out.

  12. Eddy current losses in ferromagnetic laminations

    SciTech Connect

    Serpico, C.; Visone, C.; Mayergoyz, I. D.; Basso, V.; Miano, G.

    2000-05-01

    It is demonstrated through the comparison of analytical, numerical, and experimental results that the existence of excess eddy current losses can be explained by the peculiar nature of the nonlinear diffusion of electromagnetic fields in magnetically nonlinear laminations. The essence of this peculiar nature is that nonlinear diffusion occurs as inward progress of almost rectangular profiles of magnetic flux density of variable height. Approximating actual profiles of magnetic flux density by rectangular ones, the problem of nonlinear diffusion can be treated analytically by using a simple model. The accuracy and the limit of applicability of the rectangular profile model are discussed by comparing its predictions with finite elements numerical solutions of nonlinear diffusion equation as well as with experimental results. (c) 2000 American Institute of Physics.

  13. Transient response of a laminated composite plate

    NASA Technical Reports Server (NTRS)

    Datta, S. K.; Ju, T. H.; Bratton, R. L.; Shah, A. H.

    1992-01-01

    Results are presented from an investigation of the effect of layering on transient wave propagation in a laminated cross-ply plate, giving attention to the case of 2D plane strain in the case where a line vertical force is applied on a free surface of the plate; the line may be either parallel or perpendicular to the fibers in a ply. The results are in both the time and frequency domains for the normal stress component in the x direction, at a point on the surface of the plate on which the force is applied. Comparative results are also presented for a homogeneous plate whose properties are the static effective ones, when the number of plies is large.

  14. Nuclear shape, mechanics, and mechanotransduction.

    PubMed

    Dahl, Kris Noel; Ribeiro, Alexandre J S; Lammerding, Jan

    2008-06-06

    In eukaryotic cells, the nucleus contains the genome and is the site of transcriptional regulation. The nucleus is the largest and stiffest organelle and is exposed to mechanical forces transmitted through the cytoskeleton from outside the cell and from force generation within the cell. Here, we discuss the effect of intra- and extracellular forces on nuclear shape and structure and how these force-induced changes could be implicated in nuclear mechanotransduction, ie, force-induced changes in cell signaling and gene transcription. We review mechanical studies of the nucleus and nuclear structural proteins, such as lamins. Dramatic changes in nuclear shape, organization, and stiffness are seen in cells where lamin proteins are mutated or absent, as in genetically engineered mice, RNA interference studies, or human disease. We examine the different mechanical pathways from the force-responsive cytoskeleton to the nucleus. We also highlight studies that link changes in nuclear shape with cell function during developmental, physiological, and pathological modifications. Together, these studies suggest that the nucleus itself may play an important role in the response of the cell to force.

  15. Prediction of microcracking in composite laminates under thermomechanical loading

    NASA Technical Reports Server (NTRS)

    Maddocks, Jason R.; Mcmanus, Hugh L.

    1995-01-01

    Composite laminates used in space structures are exposed to both thermal and mechanical loads. Cracks in the matrix form, changing the laminate thermoelastic properties. An analytical methodology is developed to predict microcrack density in a general laminate exposed to an arbitrary thermomechanical load history. The analysis uses a shear lag stress solution in conjunction with an energy-based cracking criterion. Experimental investigation was used to verify the analysis. Correlation between analysis and experiment is generally excellent. The analysis does not capture machining-induced cracking, or observed delayed crack initiation in a few ply groups, but these errors do not prevent the model from being a useful preliminary design tool.

  16. Accurate stress resultants equations for laminated composite deep thick shells

    SciTech Connect

    Qatu, M.S.

    1995-11-01

    This paper derives accurate equations for the normal and shear force as well as bending and twisting moment resultants for laminated composite deep, thick shells. The stress resultant equations for laminated composite thick shells are shown to be different from those of plates. This is due to the fact the stresses over the thickness of the shell have to be integrated on a trapezoidal-like shell element to obtain the stress resultants. Numerical results are obtained and showed that accurate stress resultants are needed for laminated composite deep thick shells, especially if the curvature is not spherical.

  17. Analysis of a hybrid, unidirectional buffer strip laminate

    NASA Technical Reports Server (NTRS)

    Dharani, L. R.; Goree, J. G.

    1983-01-01

    A method of analysis capable of predicting accurately the fracture behavior of a unidirectional composite laminate containing symmetrically placed buffer strips is presented. As an example, for a damaged graphite/epoxy laminate, the results demonstrate the manner in which to select the most efficient combination of buffer strip properties necessary to inhibit crack growth. Ultimate failure of the laminate after crack arrest can occur under increasing load either by continued crack extension through the buffer strips or the crack can jump the buffer strips. For some typical hybrid materials it is found that a buffer strip spacing-to-width ratio of about four to one is the most efficient.

  18. Thermal buckling of thick antisymmetric angle-ply laminates

    SciTech Connect

    Tauchert, T.R.

    1987-01-01

    The buckling behavior of moderately thick antisymmetric angle-ply laminates that are simply supported and subject to a uniform temperature rise is analyzed. Transverse shear deformation is accounted for by employing the thermoelastic version of the Reissner-Mindlin theory. Results for the classical thin-plate theory are obtained as a special case. Numerical results are presented for fiber-reinforced laminates and show the effects of ply orientation, number of layers, plate thickness, and aspect ratio on the critical buckling temperature. Finally, an optimization procedure is proposed for the design of laminates having maximum resistance to thermal buckling. 17 references.

  19. Wave propagation in graphite/epoxy laminates due to impact

    NASA Technical Reports Server (NTRS)

    Tan, T. M.; Sun, C. T.

    1982-01-01

    The low velocity impact response of graphite-epoxy laminates is investigated theoretically and experimentally. A nine-node isoparametric finite element in conjunction with an empirical contact law was used for the theoretical investigation. Flat laminates subjected to pendulum impact were used for the experimental investigation. Theoretical results are in good agreement with strain gage experimental data. The collective results of the investigation indicate that the theoretical procedure describes the impact response of the laminate up to about 150 in/sec. impact velocity.

  20. Laminated metals composites fracture and ballistic impact behavior

    SciTech Connect

    Lesuer, D.R.; Syn, C.K.; Sherby, O.D.; Wadsworth, J.

    1998-01-20

    Recent advances in the fracture and ballistic impact response of laminated metal composites (LMCs) are reviewed. The laminate structure can provide significant improvements to these properties relative to the component materials. Typical fracture and ballistic impact properties in LMCs are illustrated for systems containing Al alloys and Al matrix composites. The unique mechanisms operating in a layered structure that contribute to fracture or ballistic impact resistance are discussed. The influence of laminate architecture, component material properties and interface strength on mechanisms and properties are briefly reviewed for these Al-based LMCs.