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Sample records for joyce tombran-tink col

  1. Catachresis: Vico and Joyce.

    ERIC Educational Resources Information Center

    Levin, Samuel R.

    1987-01-01

    Considers the use and significance of catachresis in Giambattista Vico and James Joyce: in Vico's case the linguistic and metaphysical background is the one defined as actuating the utterances of the theological poets, the first mortals; and in Joyce's case it is the background of newly discovered psychological forces and their role in the conduct…

  2. Doctors, disease and James Joyce.

    PubMed

    Kaplan, Robert M

    2008-08-01

    The Irish author James Joyce is regarded as the greatest modernist writer of his time. His works, notably The Dead, A Portrait of the Artist as a Young Man, Ulysses and Finnegans Wake--are intensely autobiographic including meticulous descriptions of illness and states of health--no surprise in view of Joyce's medical history and hypochondria. The Dead revolves around the tragic love of a doomed tubercular youth. Ulysses has a graphic description of Mary Joyce-s death, a funeral and a birth; Stephen Dedalus, the character based on Joyce, attends a drinking session with medical students at the lying-in hospital just as Joyce had done as a student; references to syphilis, alcoholism and other illnesses abound. PMID:18704219

  3. The Joyce Foundation 2011 Annual Report

    ERIC Educational Resources Information Center

    O'Connell, Mary

    2012-01-01

    In a period such as this, marked by extreme political partisanship and apparent gridlock at the federal level, it would be tempting to throw up one's hands in frustration. But this is just the time that a foundation like Joyce can add the most value--because the Joyce team members are non-partisan, not subject to the pressures of political and…

  4. Developmental Writing and the Efficacy of Joyce.

    ERIC Educational Resources Information Center

    Lang, Frederick K.

    James Joyce's use of interior monologue (the interior self of the character is given directly, as though the reader were overhearing an articulation of the stream of thought and feeling flowing through the character's mind) can help basic writers in developmental classes. Students can be given excerpts from Joyce and asked to turn the sentence…

  5. James Joyce and Molly Bloom: reflections on their relationship.

    PubMed

    Jacobs, Theodore J

    2002-01-01

    The relationship between James Joyce and his memorable creation, Molly Bloom, is explored in relation to Joyce's remarkable creativity and various factors that may have contributed to it. A character forged primarily out of Joyce's perceptions of his wife Nora and memories of his mother, Molly also contains aspects of Joyce's warded-off and wished-for self-representation. A focus on both biographical and dynamic contributions to the creation of Molly helps to illuminate aspects of Joyce's psychology. PMID:12580331

  6. James Joyce: steps towards a diagnosis.

    PubMed

    Lyons, J B

    2000-12-01

    Authors whose scholarship is in the golden realm of English literature have not hesitated to make pronouncements on James Joyce's health. A publication in this genre claims he had tabes dorsalis. One feels that an authoritative comment, accepting or rejecting a diagnosis of neurosyphilis, should be provided by the Journal of the History of the Neurosciences. PMID:11232371

  7. James Joyce's "Dubliners" auf der Sekundarstufe 2 (James Joyce's "Dubliners" in Secondary Grade 2)

    ERIC Educational Resources Information Center

    Breier, Harald

    1976-01-01

    Joyce's early work offers the student an opportunity to broaden his view of and experience of the world. Some learning goals are listed, followed by methodological considerations. The experiment, done in groups, is described, as are auxiliary materials and a final test. (Text is in German.) (IFS/WGA)

  8. The Joyce of Teaching: Some Notes on Presenting James Joyce to Undergraduates.

    ERIC Educational Resources Information Center

    Scarry, John

    Problems in the teaching of James Joyce to undergraduates are explored in a discussion of the "Dubliners", "A Portrait of the Artist", "Ulysses", and "Finnegans Wake". Several multimedia approaches, including the use of records and film-making, are suggested for overcoming other problems encountered due to time factors, presentation of background…

  9. Joyce's Political Development and the Aesthetic of "Dubliners"

    ERIC Educational Resources Information Center

    Delany, Paul

    1972-01-01

    Traces James Joyce's political evolution from 1903 to his artistic maturity; suggests the chief elements of a political interpretation of Dubliners''; and argues that Joyce's political conscious derives logically from social questions posed in Dubliners.'' Comment by Gaylord C. LeRoy. (RB)

  10. Picking up Galen: James Joyce in Cecilia Street.

    PubMed

    Lyons, J B

    1997-07-01

    James Joyce (1882-1941) registered as a student of the Catholic University Medical School, Cecilia Street, in 1902. His attendance in November was brief; by early December, Joyce was in Paris. A recently-acquired Guide for Medical Students, a booklet compiled by Ambrose Birmingham, dean of the Cecilia Street school, sheds light on this hitherto obscure episode. PMID:11619448

  11. Foundation + Collaboration + Inspiration. The Joyce Foundation 2009 Annual Report

    ERIC Educational Resources Information Center

    Joyce Foundation, 2010

    2010-01-01

    Among the great strengths of a policy-oriented foundation like Joyce is the willingness to take a long view, to be patient investors in ideas that take time to have impact, and to take chances on projects that may not work out. But in times of crisis, Joyce team and partners also have an obligation to be responsive to immediate challenges in their…

  12. Joyce Carol Oates'"A Theory of Knowledge."

    ERIC Educational Resources Information Center

    Berman, Harry J.

    1993-01-01

    Presents critical reading of "A Theory of Knowledge" by Joyce Carol Oates, which appears in recently published collection of stories about aging. Analyzes story in terms of two ideas in current adult developmental theory, the idea of personal narrative as developed by Cohler, and the idea of generativity as developed by Erikson, Kotre, and…

  13. Common Core and School Librarians: An Interview with Joyce Karon

    ERIC Educational Resources Information Center

    Kramer, Pamela K.

    2011-01-01

    This article presents an interview with Joyce Karon, a former school librarian, district coordinator, and former member of the Illinois State Board of Education and the Illinois Board of Higher Education. She is currently a member of the Illinois P-20 Council and has always been a passionate advocate for school libraries. In this interview, Karon…

  14. Reading, Writing, and Psycholinguistics: An Integrated Approach Using Joyce's "Counterparts."

    ERIC Educational Resources Information Center

    Buckler, Patricia Prandini

    1985-01-01

    Explains how, in a second-semester freshman composition course, Louise Rosenblatt's model for the process of reading literature was used with James Moffett's concept of the process of writing to create a linked series of reading-writing exercises based on Joyce's "Counterparts." (FL)

  15. An Interview with ... Joyce Hinckley on the Front Line

    ERIC Educational Resources Information Center

    Morgan, Jill; Ashbaker, Betty Y.

    2004-01-01

    In this article, an interview with paraeducator Joyce Hinckley is presented. Hinckley discusses her 15-year career as a paraeducator, how she became involved with the field, and gives advice to teachers working with paraeducators. She is the mother of five children and grandmother of eight. She has lived in farming communities for most of her life.

  16. Modified 'Joyce model' of opioid dependence/withdrawal.

    PubMed

    Raffa, Robert B; Tallarida, Ronald J

    2006-12-01

    By comprehensive and detailed measurement of the time course of withdrawal signs in rats, Joyce et al. (J. Theo. Biol. 240:531-537, 2006) recently provided a creative quantitative model of the onset of drug dependence based on the requirement of protein synthesis. Because the initial model fit the data imperfectly over the full time course, those authors postulated that additional features would be needed. We report excellent fit of the data (R(2)=0.96) by adding: (1) a transient early phase, and (2) a delay in the buildup of protein. PMID:17045985

  17. Joyce Clifford the Scholar: In Her Own Words.

    PubMed

    Fulmer, Terry; Gibbons, M Patricia

    2015-01-01

    Dr Joyce H. Clifford was world renowned for her excellence in nursing administration and leadership. The purpose of this article is to examine her complete body of published scholarship and analyze her papers as a method of understanding her intellectual progression as a leader in the discipline, as well as to document how her conceptualization of professional practice and the practice environment advanced nursing practice and patient and family care. Using the qualitative method of narrative inquiry, a systematic analysis of her papers was conducted to describe the evolution of her scholarship and her impact on the discipline and patient care. We reviewed all known existing papers, categorized them into 3 stages, and discuss them here. Using quotes from her work, we have added her voice to the compelling professional practice issues she addressed in her lifetime. PMID:26517343

  18. JOYCE KILMER-SLICKROCK WILDERNESS, NORTH CAROLINA AND TENNESSEE.

    USGS Publications Warehouse

    Lesure, Frank G.; Hill, James J.

    1984-01-01

    A mineral-resource survey of the Joyce Kilmer-Slickrock Wilderness in the western part of the Blue Ridge Mountains along the border between North Carolina and Tennessee found trace amounts of such metals as copper, gold, lead, and zinc in rock, stream-sediment, and soil samples, but little promise for the occurrence of metallic-mineral resources was identified. The only apparent mineral resources are small amounts of sand and gravel and abundant rock suitable for construction materials. Oil and natural gas may be present in younger sedimentary rocks. Further seismic work and exploratory drilling are needed to evaluate the resource potential of the whole overthrust belt in this area for oil and gas.

  19. Growth of modern branched columnar stromatolites in Lake Joyce, Antarctica.

    PubMed

    Mackey, T J; Sumner, D Y; Hawes, I; Jungblut, A D; Andersen, D T

    2015-07-01

    Modern decimeter-scale columnar stromatolites from Lake Joyce, Antarctica, show a change in branching pattern during a period of lake level rise. Branching patterns correspond to a change in cyanobacterial community composition as preserved in authigenic calcite crystals. The transition in stromatolite morphology is preserved by mineralized layers that contain microfossils and cylindrical molds of cyanobacterial filaments. The molds are composed of two populations with different diameters. Large diameter molds (>2.8 μm) are abundant in calcite forming the oldest stromatolite layers, but are absent from younger layers. In contrast, <2.3 μm diameter molds are common in all stromatolites layers. Loss of large diameter molds corresponds to the transition from smooth-sided stromatolitic columns to branched and irregular columns. Mold diameters are similar to trichome diameters of the four most abundant living cyanobacteria morphotypes in Lake Joyce: Phormidium autumnale morphotypes have trichome diameters >3.5 μm, whereas Leptolyngbya antarctica, L. fragilis, and Pseudanabaena frigida morphotypes have diameters <2.3 μm. P. autumnale morphotypes were only common in mats at <12 m depth. Mats containing abundant P. autumnale morphotypes were smooth, whereas mats with few P. autumnale morphotypes contained small peaks and protruding bundles of filaments, suggesting that the absence of P. autumnale morphotypes allowed small-scale topography to develop on mats. Comparisons of living filaments and mold diameters suggest that P. autumnale morphotypes were present early in stromatolite growth, but disappeared from the community through time. We hypothesize that the mat-smoothing behavior of P. autumnale morphotypes inhibited nucleation of stromatolite branches. When P. autumnale morphotypes were excluded from the community, potentially reflecting a rise in lake level, short-wavelength roughness provided nuclei for stromatolite branches. This growth history provides a

  20. Uveitic secondary glaucoma: influence in James Joyce's (1882-1941) last works.

    PubMed

    Ascaso, Francisco J; Bosch, Jordi

    2010-02-01

    James Joyce, considered one of the pre-eminent novelists of the 20th century, attained international renown with his work Ulysses. Its lack of standard punctuation makes it difficult to read. An example would be the famous non-punctuated 'Molly Bloom soliloquy' in the last chapter of Ulysses. Why is Joyce considered so difficult to read? He wrote and proofread Ulysses and Finnegans wake, his last works, during his battle against glaucoma, when his vision was seriously blurred. The distracting and confusing diacritical marks might be explained by Joyce's reduced visual acuity. Could Ulysses and Finnegans wake have been different if Joyce's visual problems had begun in the second rather than the first half of 20th century? PMID:20207905

  1. Joyce After Flaubert: the cuckold as imperfect physician, the writer as physiologist.

    PubMed

    Bénéjam, Valérie

    2008-01-01

    Although Joyce was not as familiar with the practice and theory of medicine as was Gustave Flaubert, this article argues that, through Flaubert's legacy, Joyce's writing was influenced by the French school of medical thought. Several aspects of Flaubert's style and narration-what has been dubbed his "medical realism"-were taken up by Joyce: the artist's impersonal perspective, the precision of descriptions, and the materialist attack against Romanticism, as well as the irony built into the narrative voice through free indirect discourse. While the cuckold in Madame Bovary is an incompetent surgeon serving as foil to the precise description of sentiments offered by the narrator, Joyce's cuckold in Ulysses is an amateur physiologist, both perspicacious and sympathetic to human suffering. Bloom's interest in internal bodily processes opens up new dimensions for a modernist aesthetics as he relates physiology and psychology, in accordance with the theories of Xavier Bichat, Etienne Bonnot de Condillac, and Pierre Jean Georges Cabanis. In keeping with such focus, Joyce's physiological version of stream-of-consciousness stems from Flaubert's clinical description of characters, but he directs matters even further inward. PMID:20836269

  2. Coincidence, historical repetition, and self-knowledge: Jung, Vico, and Joyce.

    PubMed

    Verene, Donald Phillip

    2002-07-01

    Jung develops synchronicity as an a causal principle of connection by recounting various examples of meaningful coincidence from experience and by analysing various systems of divination, notably the I Ching. Philosophical theory of causality has given no significant attention to synchronicity; the events of synchronicity are regarded as chance. The Neapolitan philosopher Giambattista Vico (1668-1744) developed a doctrine of historical experience and of self-knowledge that grounds the phenomenon of synchronicity in a metaphysics. James Joyce employed Vico's conception of language and historical cycles as the basis of Joyce's final literary work, Finnegans Wake. Vico's metaphysical sense of synchronicity and Joyce's literary formulation offer a grounding of this principle in non-divinatory sources in modern Western thought, something which Jung's discussion does not provide. These philosophical and literary perspectives complement Jung's to offer an expanded context in which to recognize synchronicity and to make sense of it. PMID:12174547

  3. Us and Them: Joyce Carol Oates and the Stories Students Tell

    ERIC Educational Resources Information Center

    DeGenaro, William

    2007-01-01

    Responding with strategic empathy to the traumatic stories students share with us provides an opportunity to break down an elitist binary between teacher and student. Joyce Carol Oates's novel "them" can serve as a cautionary tale for understanding the dangers of disregarding student trauma. (Contains 2 notes.)

  4. Further Evidence that Legalized Abortion Lowered Crime: A Reply to Joyce

    ERIC Educational Resources Information Center

    Donohue, John J., III; Levitt, Steven D.

    2004-01-01

    Joyce's failure to uncover a negative relationship between crime and abortion was because of his decision to concentrate on a non-representative six-year period. Evidence supporting the claims that the crack-cocaine epidemic hit the high-abortion early-legalizing states earlier and more severely than other states of the U.S in 1970 is presented.

  5. [Portrait of the artist as a sick man. Rheumatological pathography of James Joyce (1882-1941)].

    PubMed

    Ventura, L

    2008-01-01

    James Joyce, unanimously considered one of the greatest novelists of the 20th century, suffered from several diseases. A series of adverse circumstances progressively deteriorated his health, already precarious because of his very disorderly life habits. Aim of the present study is to summarize the various organic diseases Joyce suffered during his lifetime, as long as the main diagnostic conclusions found in scientific literature. Severe eye problems, caused by recurrent iritis attacks even complicated by glaucoma and cataracts, led him almost to blindness. Undernourishment and irregular eating, great anxiety and alcohol abuse were the major causes of the peptic ulcer which tortured him for many years, causing his final death. To these conditions should also be added dental caries, venereal diseases and recurrent polyarthritis. The hypothesis according which Joyce suffered from neurosyphilis is still debated and should be sufficiently demonstrated, whereas a spondyloarthropathy, either Reiter's syndrome or ankylosing spondylitis, appears more likely. Therapies against these diseases, easily treated today, did not result efficient because of his poor compliance, as well as the state of the art of medical science during his lifetime. A detailed paleopathologic study of Joyce's human remains could allow to solve the diagnostic doubts concerning his main disease. PMID:18651061

  6. Surviving the "School of Slavery": Acculturation in Sharon Draper's "Copper Sun" and Joyce Hansen's "The Captive"

    ERIC Educational Resources Information Center

    Chandler, Karen Michele

    2016-01-01

    Although children's literature has long alluded to cultural connections between Africans and African Americans, very few texts establish clear lines of influence between particular African ethnic groups and African American characters and communities. Joyce Hansen's "The Captive" (1994) and Sharon Draper's "Copper Sun" (2006)…

  7. Reflections on Differentiation: An Interview with Joyce VanTassel-Baska

    ERIC Educational Resources Information Center

    Understanding Our Gifted, 2007

    2007-01-01

    Joyce VanTassel-Baska is the Executive Director of the Center for Gifted Education and the Jody and Layton Smith Professor of Education at the College of William & Mary, Williamsburg, Virginia. For the past 20 years, the Integrated Curriculum Model, based on research of what works with gifted learners, has been the foundation of curriculum…

  8. ‘Bellography’: Life and Contributions of Ross and Joyce Bell, two New England Naturalists

    PubMed Central

    Spence, John R.; Ball, George E.; Davidson, Robert L.; Rykken, Jessica J.

    2011-01-01

    Abstract The lives and contributions of Ross and Joyce Bell are described with particular attention to studies of invertebrate natural history in the state of Vermont and carabid beetles of several groups, including the world rhysodine fauna. Their work, all done at the University of Vermont, was mainly taxonomic in nature and included aspects of the biology of the species considered. During their careers they described more than 75% of the c. 340 rhysodine species known to science. Ross Bell also wrote a number of seminal papers about the basal relationships of the Adephaga and the comparative anatomy of carabid coxal cavities. Ross and Joyce inspired several generations of students at UVM to take up advanced work in entomology and natural history. PMID:22371660

  9. Joyce and Ulysses: integrated and user-friendly tools for the parameterization of intramolecular force fields from quantum mechanical data.

    PubMed

    Barone, Vincenzo; Cacelli, Ivo; De Mitri, Nicola; Licari, Daniele; Monti, Susanna; Prampolini, Giacomo

    2013-03-21

    The Joyce program is augmented with several new features, including the user friendly Ulysses GUI, the possibility of complete excited state parameterization and a more flexible treatment of the force field electrostatic terms. A first validation is achieved by successfully comparing results obtained with Joyce2.0 to literature ones, obtained for the same set of benchmark molecules. The parameterization protocol is also applied to two other larger molecules, namely nicotine and a coumarin based dye. In the former case, the parameterized force field is employed in molecular dynamics simulations of solvated nicotine, and the solute conformational distribution at room temperature is discussed. Force fields parameterized with Joyce2.0, for both the dye's ground and first excited electronic states, are validated through the calculation of absorption and emission vertical energies with molecular mechanics optimized structures. Finally, the newly implemented procedure to handle polarizable force fields is discussed and applied to the pyrimidine molecule as a test case. PMID:23389748

  10. Erich Fromm's productivity: creativity as exemplified by Joyce's blooming of Leopold and Molly.

    PubMed

    Harrell, Valentina

    2005-01-01

    According to Erich Fromm, the productive character expresses what he called the life force in ways that are, by nature, artistic in quality. His valuing of the vital and the artistic, fueled by this life force in our potential to be human, serves as the cornerstone of his model of psychoanalysis. Gilbert Rose, author of Necessary Illusion: Art as "Witness", also has identified the parallel between analytic and artistic processes, building on the assumption that the therapeutic alliance and the aesthetic alliance are one and the same. In elaborating this assumption and extending the parallel between Rose and Fromm, I draw the conclusion that the qualities of relationships have an impact on one's ability to express oneself creatively, to live an artistic life--that is, to live productively. Psychoanalysis is, in practice, a process that is artistic, creative, and re-creative in nature. To the degree that authentic expression of emotionally charged implicit knowledge of the ineffable (the emotional life that resides deep in the bodymind) results in transformation and healing, the process is artistic in nature. This process is exemplified by examining the life and literary creations of James Joyce, especially Joyce's characters of Molly and Leopold Bloom in Ulysses. PMID:15953782

  11. An Addendum to Leading Change in Gifted Education: The Festschrift of Dr. Joyce VanTassel-Baska

    ERIC Educational Resources Information Center

    Lord, E. Wayne; Swanson, Julie Dingle; Breard, Nan; Drain, Denise; Thomas, Kianga R.; Thomas, Steve; Dolph, Katie A.; Price, R. Douglas

    2009-01-01

    On March 13, 2009, Dr. Joyce VanTassel-Baska, an eminent leader in gifted education, was presented with a Festschrift, a volume of articles submitted by colleagues as well as former and current graduate students to celebrate her life's work and influence on the field. This volume is a free supplement from The College of William and Mary's Center…

  12. Synergistic observations of cloud properties and their related uncertainties at JOYCE

    NASA Astrophysics Data System (ADS)

    Ebell, K.; Hünerbein, A.; Orlandi, E.; Löhnert, U.; Crewell, S.

    2013-12-01

    In order to improve the representation of clouds and the associated processes in numerical weather prediction and climate models, accurate observations of the macro- and microphysical properties of clouds are essential. However, depending on the retrieval algorithm applied, the results may differ substantially (e.g. Huang et al., 2012). In order to track down these differences, a 'blindtest' experiment of four different liquid water cloud retrieval algorithms has been carried out within the European COST action EG-CLIMET. The need to better quantify and understand the uncertainties related to cloud retrievals has also been a key issue in recent scientific collaborations: e.g. the objective of the EU/DOE ground-based cloud and precipitation retrieval workshop held in Cologne in May 2013 was to advance algorithm development and uncertainty quantification for retrieving cloud and precipitation properties from ground-based remote sensors through international scientific collaboration and data sharing. In this contribution, we present a 1D-VAR retrieval algorithm of microphysical properties of clouds, which integrates the information of various ground-based remote sensing instruments and provides a physically consistent picture of clouds. The use of a variational approach allows to properly assess the uncertainties due to measurement error, forward model and prior information. We will discuss the influence of the different uncertainty sources on the solution and characterize the information content from the different sensors. The retrieval method is applied to measurements from the Jülich Observatory for Cloud Evolution (JOYCE) located at the Research Center Jülich in Germany. The core instruments of JOYCE for deriving cloud properties include cloud radar, ceilometer, Doppler lidar, a 14-channel microwave radiometer (MWR) and an infrared spectrometer. The information from the SEVIRI instrument on board the MSG satellite can be further used to constrain the solution

  13. Chaos Theory and James Joyce's "ulysses": Leopold Bloom as a Human COMPLEX@SYSTEM^

    NASA Astrophysics Data System (ADS)

    Mackey, Peter Francis

    1995-01-01

    These four ideas apply as much to our lives as to the life of Leopold Bloom: (1) A trivial decision can wholly change a life. (2) A chance encounter can dramatically alter life's course. (3) A contingent nexus exists between consciousness and environment. (4) A structure of meaning helps us interpret life's chaos. These ideas also relate to a contemporary science called by some "chaos theory." The connection between Ulysses and chaos theory enhances our understanding of Bloom's day; it also suggests that this novel may be about the real process of life itself. The first chapter explains how Joyce's own essays and comments to friends compel attention to the links between Ulysses and chaos theory. His scientific contemporaries anticipated chaos theory, and their ideas seem to have rubbed off on him. We see this in his sense of trivial things and chance, his modernistic organizational impulses, and the contingent nature of Bloom's experience. The second chapter studies what chaos theory and Joyce's ideas tell us about "Ithaca," the episode which particularly implicates our processes of interpreting this text as well as life itself as we face their chaos. The third chapter examines Bloom's close feel for the aboriginal world, a contingency that clarifies his vulnerability to trivial changes. The fourth chapter studies how Bloom's stream of consciousness unfolds--from his chance encounters with trivial things. Beneath this stream's seeming chaos, Bloom's distinct personality endures, similar to how Joyce's schemas give Ulysses an imbedded, underlying order. The fifth chapter examines how trivial perturbations, such as Lyons' misunderstanding about "Throwaway," produce small crises for Bloom, exacerbating his seeming impotence before his lonely "fate.". The final chapter analyzes Bloom's views that fate and chance dictate his life. His views provide an opportunity to explore the implications chaos theory has for our understanding of free will and determinism. Ultimately

  14. COL4A1 and COL4A2 mutations and disease: insights into pathogenic mechanisms and potential therapeutic targets

    PubMed Central

    Kuo, Debbie S.; Labelle-Dumais, Cassandre; Gould, Douglas B.

    2012-01-01

    Heterotrimers composed of collagen type IV alpha 1 (COL4A1) and alpha 2 (COL4A2) constitute one of the most abundant components of nearly all basement membranes. Accordingly, mutations in COL4A1 or COL4A2 are pleiotropic and contribute to a broad spectrum of disorders, including myopathy, glaucoma and hemorrhagic stroke. Here, we summarize the contributions of COL4A1 and COL4A2 mutations in human disease, integrate knowledge gained from model organisms and evaluate the implications for pathogenic mechanisms and therapeutic approaches. PMID:22914737

  15. MUSCLE: MUltiscale Spherical-ColLapse Evolution

    NASA Astrophysics Data System (ADS)

    Neyrinck, Mark C.

    2016-05-01

    MUSCLE (MUltiscale Spherical ColLapse Evolution) produces low-redshift approximate N-body realizations accurate to few-Megaparsec scales. It applies a spherical-collapse prescription on multiple Gaussian-smoothed scales. It achieves higher accuracy than perturbative schemes (Zel'dovich and second-order Lagrangian perturbation theory - 2LPT), and by including the void-in-cloud process (voids in large-scale collapsing regions), solves problems with a single-scale spherical-collapse scheme.

  16. Multicanonical simulation of the Domb-Joyce model and the Gō model: new enumeration methods for self-avoiding walks

    NASA Astrophysics Data System (ADS)

    Shirai, Nobu C.; Kikuchi, Macoto

    2013-08-01

    We develop statistical enumeration methods for self-avoiding walks using a powerful sampling technique called the multicanonical Monte Carlo method. Using these methods, we estimate the numbers of the two dimensional N-step self-avoiding walks up to N = 256 with statistical errors. The developed methods are based on statistical mechanical models of paths which include self-avoiding walks. The criterion for selecting a suitable model for enumerating self-avoiding walks is whether or not the configuration space of the model includes a set for which the number of the elements can be exactly counted. We call this set a scale fixing set. We selected the following two models which satisfy the criterion: the Gō model for lattice proteins and the Domb-Joyce model for generalized random walks. There is a contrast between these two models in the structures of the configuration space. The configuration space of the Gō model is defined as the universal set of self-avoiding walks, and the set of the ground state conformation provides a scale fixing set. On the other hand, the configuration space of the Domb-Joyce model is defined as the universal set of random walks which can be used as a scale fixing set, and the set of the ground state conformation is the same as the universal set of self-avoiding walks. From the perspective of enumeration performance, we conclude that the Domb-Joyce model is the better of the two. The reason for the performance difference is partly explained by the existence of the first-order phase transition of the Gō model.

  17. Rare hereditary COL4A3/COL4A4 variants may be mistaken for familial focal segmental glomerulosclerosis

    PubMed Central

    Malone, Andrew F; Phelan, Paul J; Hall, Gentzon; Cetincelik, Umran; Homstad, Alison; Alonso, Andrea; Jiang, Ruiji; Lindsey, Thomas; Wu, Guanghong; Sparks, Matthew A; Smith, Stephen R; Webb, Nicholas J A; Kalra, Philip; Adeyemo, Adebowale; Shaw, Andrey S; Conlon, Peter J; Jennette, J Charles; Howell, David N; Winn, Michelle P; Gbadegesin, Rasheed A

    2014-01-01

    Focal segmental glomerulosclerosis (FSGS) is a histological lesion with many causes including inherited genetic defects with significant proteinuria being the predominant clinical finding at presentation. Mutations in COL4A3 and COL4A4 are known to cause Alport syndrome, thin basement membrane nephropathy, and to result in pathognomonic glomerular basement membrane findings. Secondary FSGS is known to develop in classic Alport Syndrome at later stages of the disease. Here, we present seven families with rare or novel variants in COL4A3 or COL4A4 (six with single and one with two heterozygous variants) from a cohort of 70 families with a diagnosis of hereditary FSGS. The predominant clinical findings at diagnosis were proteinuria associated with hematuria. In all seven families, there were individuals with nephrotic range proteinuria with histologic features of FSGS by light microscopy. In one family, electron microscopy showed thin glomerular basement membrane, but four other families had variable findings inconsistent with classical Alport nephritis. There was no recurrence of disease after kidney transplantation. Families with COL4A3 and COL4A4 variants that segregated with disease represent 10% of our cohort. Thus, COL4A3 and COL4A4 variants should be considered in the interpretation of next-generation sequencing data from such patients. Furthermore, this study illustrates the power of molecular genetic diagnostics in the clarification of renal phenotypes. PMID:25229338

  18. Engaging Families in Partnership Programs to Promote Student Success: Q&A for Dr. Joyce L. Epstein. REL Mid-Atlantic Educator Effectiveness Webinar Series

    ERIC Educational Resources Information Center

    Regional Educational Laboratory Mid-Atlantic, 2015

    2015-01-01

    In this webinar, Dr. Joyce Epstein, Director of the Center on School, Family, and Community Partnerships and the National Network of Partnership Schools, discussed what the research says about effective family engagement. The webinar and PowerPoint presentation are also available. A brief list of resources is included.

  19. COL4A1 Mutation in Preterm Intraventricular Hemorrhage

    PubMed Central

    Bilguvar, Kaya; DiLuna, Michael L.; Bizzarro, Matthew J.; Bayri, Yasar; Schneider, Karen C.; Lifton, Richard P.; Gunel, Murat; Ment, Laura R.

    2010-01-01

    Intraventricular hemorrhage is a common complication of preterm infants. Mutations in the type IV procollagen gene, COL4A1, are associated with cerebral small vessel disease with hemorrhage in adults and fetuses. We report a rare variant in COL4A1 associated with intraventricular hemorrhage in dizygotic preterm twins. These results expand the spectrum of diseases attributable to mutations in type IV procollagens. PMID:19840616

  20. Developmental consequences of the ColQ/MuSK interactions.

    PubMed

    Karmouch, Jennifer; Dobbertin, Alexandre; Sigoillot, Severine; Legay, Claire

    2013-03-25

    CollagenQ (ColQ) is a specific collagen that anchors acetylcholinesterase (AChE) in the synaptic basal lamina of the neuromuscular junction (NMJ). Over 30 mutations in the COLQ gene have been identified that are responsible for a congenital myasthenic syndrome with AChE deficiency, highlighting the importance of this collagen in the physiology of the NMJ. The anchoring of AChE at the synapse requires the interaction of ColQ with MuSK (Muscle-Specific Kinase), a tyrosine kinase expressed on the muscle membrane that is necessary for the formation and the maintenance of the NMJ. MuSK forms with its co-receptor LRP4, a member of the Low-density Related Protein family, a receptor complex for agrin and Wnts, representing the core system from which the postsynaptic domain is built, the growth cone attracted and the presynaptic element instructed for some aspects of its differentiation. Therefore, the discovery that ColQ binds to MuSK prompted us to study a possible regulatory function of ColQ during NMJ development. In this review, after a brief survey on ColQ, we summarize our recent data demonstrating that ColQ, in addition to its anchoring role, exerts signaling functions and controls some aspects of postsynaptic differentiation such as the clustering of acetylcholine receptors. Our results also strengthen the hypothesis that the defects observed in synaptic congenital myasthenic syndromes might be linked, at least in part, to alterations of ColQ signaling functions and not only to AChE deficiency. Finally, we discuss future research directions to understand how ColQ may modulate the action of the other ligands of the MuSK/LRP4 complex and cooperate with them to coordinate the different steps of NMJ formation and maintenance. PMID:23089045

  1. Expression of colSR Genes Increased in the rpf Mutants of Xanthomonas oryzae pv. oryzae KACC10859

    PubMed Central

    Noh, Young-Hee; Kim, Sun-Young; Han, Jong-Woo; Seo, Young-Su; Cha, Jae-Soon

    2014-01-01

    The rpf genes and colSXOO1207/colRXOO1208 were known to require for virulence of Xanthomonas oryzae pv. oryzae (Xoo). In Xoo KACC10331 genome, two more colS/colR genes, colSXOO3534 (raxH)/colRXOO3535 (raxR) and colSXOO3762/colRXOO3763 were annotated. The colSXOO3534/colRXOO3535 were known to control AvrXa21 activity and functions of colSXOO3762/colRXOO3763 were unknown in Xoo. To characterize the relationship between rpf and colS/colR genes, expression of colS/colR genes in Rpf mutants of Xoo were analyzed with quantitative reverse transcription PCR (qRT-PCR). Expressions of all three colS/colR genes increased in the rpfF mutant in which DSF synthesis is defective. Expression of colSXOO1207/colRXOO1208, colSXOO3534/colRXOO3535 and colSXOO3762/colRXOO3763 increased 2, 2–7, 3–13 folds respectively. Expression of colSXOO3534 and colSXOO3762 also increased 2–4 folds in the rpfG mutant in which the signal from DSF is no longer transferred to down-stream. Expression of the other colS/colR genes was not significantly changed in the rpfG mutant compared to the wild type. Since RpfF and RpfG are responsible for DSF synthesis and signal transfer from DSF to down-stream to regulate virulence gene expression, these results suggest that the DSF and DSF-mediated signal regulate negatively three colS/colR genes in Xoo. PMID:25289017

  2. Complete DNA Sequence of a ColBM Plasmid from Avian Pathogenic Escherichia coli Suggests that It Evolved from Closely Related ColV Virulence Plasmids†

    PubMed Central

    Johnson, Timothy J.; Johnson, Sara J.; Nolan, Lisa K.

    2006-01-01

    Avian pathogenic Escherichia coli (APEC), an extraintestinal pathogenic E. coli causing colibacillosis in birds, is responsible for significant economic losses for the poultry industry. Recently, we reported that the APEC pathotype was characterized by possession of a set of genes contained within a 94-kb cluster linked to a ColV plasmid, pAPEC-O2-ColV. These included sitABCD, genes of the aerobactin operon, hlyF, iss, genes of the salmochelin operon, and the 5′ end of cvaB of the ColV operon. However, the results of gene prevalence studies performed among APEC isolates revealed that these traits were not always linked to ColV plasmids. Here, we present the complete sequence of a 174-kb plasmid, pAPEC-O1-ColBM, which contains a putative virulence cluster similar to that of pAPEC-O2-ColV. These two F-type plasmids share remarkable similarity, except that they encode the production of different colicins; pAPEC-O2-ColV contains an intact ColV operon, and pAPEC-O1-ColBM encodes the colicins B and M. Interestingly, remnants of the ColV operon exist in pAPEC-O1-ColBM, hinting that ColBM-type plasmids may have evolved from ColV plasmids. Among APEC isolates, the prevalence of ColBM sequences helps account for the previously observed differences in prevalence between genes of the “conserved” portion of the putative virulence cluster of pAPEC-O2-ColV and those genes within its “variable” portion. These results, in conjunction with Southern blotting and probing of representative ColBM-positive strains, indicate that this “conserved” cluster of putative virulence genes is primarily linked to F-type virulence plasmids among the APEC isolates studied. PMID:16885466

  3. Cloning, Purification and Characterization of the Collagenase ColA Expressed by Bacillus cereus ATCC 14579.

    PubMed

    Abfalter, Carmen M; Schönauer, Esther; Ponnuraj, Karthe; Huemer, Markus; Gadermaier, Gabriele; Regl, Christof; Briza, Peter; Ferreira, Fatima; Huber, Christian G; Brandstetter, Hans; Posselt, Gernot; Wessler, Silja

    2016-01-01

    Bacterial collagenases differ considerably in their structure and functions. The collagenases ColH and ColG from Clostridium histolyticum and ColA expressed by Clostridium perfringens are well-characterized collagenases that cleave triple-helical collagen, which were therefore termed as ´true´ collagenases. ColA from Bacillus cereus (B. cereus) has been added to the collection of true collagenases. However, the molecular characteristics of B. cereus ColA are less understood. In this study, we identified ColA as a secreted true collagenase from B. cereus ATCC 14579, which is transcriptionally controlled by the regulon phospholipase C regulator (PlcR). B. cereus ATCC 14579 ColA was cloned to express recombinant wildtype ColA (ColAwt) and mutated to a proteolytically inactive (ColAE501A) version. Recombinant ColAwt was tested for gelatinolytic and collagenolytic activities and ColAE501A was used for the production of a polyclonal anti-ColA antibody. Comparison of ColAwt activity with homologous proteases in additional strains of B. cereus sensu lato (B. cereus s.l.) and related clostridial collagenases revealed that B. cereus ATCC 14579 ColA is a highly active peptidolytic and collagenolytic protease. These findings could lead to a deeper insight into the function and mechanism of bacterial collagenases which are used in medical and biotechnological applications. PMID:27588686

  4. Structures of three polycystic kidney disease-like domains from Clostridium histolyticum collagenases ColG and ColH.

    PubMed

    Bauer, Ryan; Janowska, Katarzyna; Taylor, Kelly; Jordan, Brad; Gann, Steve; Janowski, Tomasz; Latimer, Ethan C; Matsushita, Osamu; Sakon, Joshua

    2015-03-01

    Clostridium histolyticum collagenases ColG and ColH are segmental enzymes that are thought to be activated by Ca(2+)-triggered domain reorientation to cause extensive tissue destruction. The collagenases consist of a collagenase module (s1), a variable number of polycystic kidney disease-like (PKD-like) domains (s2a and s2b in ColH and s2 in ColG) and a variable number of collagen-binding domains (s3 in ColH and s3a and s3b in ColG). The X-ray crystal structures of Ca(2+)-bound holo s2b (1.4 Å resolution, R = 15.0%, Rfree = 19.1%) and holo s2a (1.9 Å resolution, R = 16.3%, Rfree = 20.7%), as well as of Ca(2+)-free apo s2a (1.8 Å resolution, R = 20.7%, Rfree = 27.2%) and two new forms of N-terminally truncated apo s2 (1.4 Å resolution, R = 16.9%, Rfree = 21.2%; 1.6 Å resolution, R = 16.2%, Rfree = 19.2%), are reported. The structurally similar PKD-like domains resemble the V-set Ig fold. In addition to a conserved β-bulge, the PKD-like domains feature a second bulge that also changes the allegiance of the subsequent β-strand. This β-bulge and the genesis of a Ca(2+) pocket in the archaeal PKD-like domain suggest a close kinship between bacterial and archaeal PKD-like domains. Different surface properties and indications of different dynamics suggest unique roles for the PKD-like domains in ColG and in ColH. Surface aromatic residues found on ColH s2a-s2b, but not on ColG s2, may provide the weak interaction in the biphasic collagen-binding mode previously found in s2b-s3. B-factor analyses suggest that in the presence of Ca(2+) the midsection of s2 becomes more flexible but the midsections of s2a and s2b stay rigid. The different surface properties and dynamics of the domains suggest that the PKD-like domains of M9B bacterial collagenase can be grouped into either a ColG subset or a ColH subset. The conserved properties of PKD-like domains in ColG and in ColH include Ca(2+) binding. Conserved residues not only interact with Ca(2+), but also

  5. Coronary-Heart-Disease-Associated Genetic Variant at the COL4A1/COL4A2 Locus Affects COL4A1/COL4A2 Expression, Vascular Cell Survival, Atherosclerotic Plaque Stability and Risk of Myocardial Infarction

    PubMed Central

    Pu, Xiangyuan; Ren, Meixia; An, Weiwei; Zhang, Ruoxin; Yan, Shunying; Situ, Haiteng; He, Xinjie; Chen, Yequn; Tan, Xuerui; Xiao, Qingzhong; Tucker, Arthur T.; Caulfield, Mark J.; Ye, Shu

    2016-01-01

    Genome-wide association studies have revealed an association between coronary heart disease (CHD) and genetic variation on chromosome 13q34, with the lead single nucleotide polymorphism rs4773144 residing in the COL4A2 gene in this genomic region. We investigated the functional effects of this genetic variant. Analyses of primary cultures of vascular smooth muscle cells (SMCs) and endothelial cells (ECs) from different individuals showed a difference between rs4773144 genotypes in COL4A2 and COL4A1 expression levels, being lowest in the G/G genotype, intermediate in A/G and highest in A/A. Chromatin immunoprecipitation followed by allelic imbalance assays of primary cultures of SMCs and ECs that were of the A/G genotype revealed that the G allele had lower transcriptional activity than the A allele. Electrophoretic mobility shift assays and luciferase reporter gene assays showed that a short DNA sequence encompassing the rs4773144 site interacted with a nuclear protein, with lower efficiency for the G allele, and that the G allele sequence had lower activity in driving reporter gene expression. Analyses of cultured SMCs from different individuals demonstrated that cells of the G/G genotype had higher apoptosis rates. Immunohistochemical and histological examinations of ex vivo atherosclerotic coronary arteries from different individuals disclosed that atherosclerotic plaques with the G/G genotype had lower collagen IV abundance and thinner fibrous cap, a hallmark of unstable, rupture-prone plaques. A study of a cohort of patients with angiographically documented coronary artery disease showed that patients of the G/G genotype had higher rates of myocardial infarction, a phenotype often caused by plaque rupture. These results indicate that the CHD-related genetic variant at the COL4A2 locus affects COL4A2/COL4A1 expression, SMC survival, and atherosclerotic plaque stability, providing a mechanistic explanation for the association between the genetic variant and CHD

  6. Coronary-Heart-Disease-Associated Genetic Variant at the COL4A1/COL4A2 Locus Affects COL4A1/COL4A2 Expression, Vascular Cell Survival, Atherosclerotic Plaque Stability and Risk of Myocardial Infarction.

    PubMed

    Yang, Wei; Ng, Fu Liang; Chan, Kenneth; Pu, Xiangyuan; Poston, Robin N; Ren, Meixia; An, Weiwei; Zhang, Ruoxin; Wu, Jingchun; Yan, Shunying; Situ, Haiteng; He, Xinjie; Chen, Yequn; Tan, Xuerui; Xiao, Qingzhong; Tucker, Arthur T; Caulfield, Mark J; Ye, Shu

    2016-07-01

    Genome-wide association studies have revealed an association between coronary heart disease (CHD) and genetic variation on chromosome 13q34, with the lead single nucleotide polymorphism rs4773144 residing in the COL4A2 gene in this genomic region. We investigated the functional effects of this genetic variant. Analyses of primary cultures of vascular smooth muscle cells (SMCs) and endothelial cells (ECs) from different individuals showed a difference between rs4773144 genotypes in COL4A2 and COL4A1 expression levels, being lowest in the G/G genotype, intermediate in A/G and highest in A/A. Chromatin immunoprecipitation followed by allelic imbalance assays of primary cultures of SMCs and ECs that were of the A/G genotype revealed that the G allele had lower transcriptional activity than the A allele. Electrophoretic mobility shift assays and luciferase reporter gene assays showed that a short DNA sequence encompassing the rs4773144 site interacted with a nuclear protein, with lower efficiency for the G allele, and that the G allele sequence had lower activity in driving reporter gene expression. Analyses of cultured SMCs from different individuals demonstrated that cells of the G/G genotype had higher apoptosis rates. Immunohistochemical and histological examinations of ex vivo atherosclerotic coronary arteries from different individuals disclosed that atherosclerotic plaques with the G/G genotype had lower collagen IV abundance and thinner fibrous cap, a hallmark of unstable, rupture-prone plaques. A study of a cohort of patients with angiographically documented coronary artery disease showed that patients of the G/G genotype had higher rates of myocardial infarction, a phenotype often caused by plaque rupture. These results indicate that the CHD-related genetic variant at the COL4A2 locus affects COL4A2/COL4A1 expression, SMC survival, and atherosclerotic plaque stability, providing a mechanistic explanation for the association between the genetic variant and CHD

  7. Collagen type IV-related nephropathies in Portugal: pathogenic COL4A3 and COL4A4 mutations and clinical characterization of 25 families.

    PubMed

    Nabais Sá, M J; Storey, H; Flinter, F; Nagel, M; Sampaio, S; Castro, R; Araújo, J A; Gaspar, M A; Soares, C; Oliveira, A; Henriques, A C; da Costa, A G; Abreu, C P; Ponce, P; Alves, R; Pinho, L; Silva, S E; de Moura, C P; Mendonça, L; Carvalho, F; Pestana, M; Alves, S; Carvalho, F; Oliveira, J P

    2015-11-01

    Pathogenic mutations in genes COL4A3/COL4A4 are responsible for autosomal Alport syndrome (AS) and thin basement membrane nephropathy (TBMN). We used Sanger sequencing to analyze all exons and splice site regions of COL4A3/COL4A4, in 40 unrelated Portuguese probands with clinical suspicion of AS/TBMN. To assess genotype-phenotype correlations, we compared clinically relevant phenotypes/outcomes between homozygous/compound heterozygous and apparently heterozygous patients. Seventeen novel and four reportedly pathogenic COL4A3/COL4A4 mutations were identified in 62.5% (25/40) of the probands. Regardless of the mutated gene, all patients with ARAS manifested chronic renal failure (CRF) and hearing loss, whereas a minority of the apparently heterozygous patients had CRF or extrarenal symptoms. CRF was diagnosed at a significantly younger age in patients with ARAS. In our families, the occurrence of COL4A3/COL4A4 mutations was higher, while the prevalence of XLAS was lower than expected. Overall, a pathogenic COL4A3/COL4A4/COL4A5 mutation was identified in >50% of patients with fewer than three of the standard diagnostic criteria of AS. With such a population background, simultaneous next-generation sequencing of all three genes may be recommended as the most expedite approach to diagnose collagen IV-related glomerular basement membrane nephropathies. PMID:25307543

  8. Dietary Lecithin Decreases Skeletal Muscle COL1A1 and COL3A1 Gene Expression in Finisher Gilts

    PubMed Central

    Akit, Henny; Collins, Cherie; Fahri, Fahri; Hung, Alex; D’Souza, Daryl; Leury, Brian; Dunshea, Frank

    2016-01-01

    Simple Summary In this study, the effect of dietary lecithin on skeletal muscle gene expression of collagen precursors and enzymes was investigated in gilts. Thirty-six finisher gilts were fed with diets containing either 0, 4, 20 or 80 g/kg soybean lecithin for six weeks. Then, rectus abdominis muscle was sampled and analyzed for eight genes involved in collagen synthesis and degradation (COL1A1, COL3A1, MMP-1, MMP-13, TIMP-1, TIMP-3, lysyl oxidase and α-subunit P4H) using quantitative real-time PCR. The results showed that lecithin down-regulated COL1A1 and COL3A1 as well as tended to down-regulate α-subunit P4H expression. Abstract The purpose of this study was to investigate the effect of dietary lecithin on skeletal muscle gene expression of collagen precursors and enzymes involved in collagen synthesis and degradation. Finisher gilts with an average start weight of 55.9 ± 2.22 kg were fed diets containing either 0, 4, 20 or 80 g/kg soybean lecithin prior to harvest for six weeks and the rectus abdominis muscle gene expression profile was analyzed by quantitative real-time PCR. Lecithin treatment down-regulated Type I (α1) procollagen (COL1A1) and Type III (α1) procollagen (COL3A1) mRNA expression (p < 0.05, respectively), indicating a decrease in the precursors for collagen synthesis. The α-subunit of prolyl 4-hydroxylase (P4H) mRNA expression also tended to be down-regulated (p = 0.056), indicating a decrease in collagen synthesis. Decreased matrix metalloproteinase-1 (MMP-1) mRNA expression may reflect a positive regulatory response to the reduced collagen synthesis in muscle from the pigs fed lecithin (p = 0.035). Lecithin had no effect on tissue inhibitor metalloproteinase-1 (TIMP-1), matrix metalloproteinase-13 (MMP-13) and lysyl oxidase mRNA expression. In conclusion, lecithin down-regulated COL1A1 and COL3A1 as well as tended to down-regulate α-subunit P4H expression. However, determination of muscle collagen content and solubility are required

  9. Genetics Home Reference: COL4A1-related brain small-vessel disease

    MedlinePlus

    ... COL4A1-related brain small-vessel disease COL4A1-related brain small-vessel disease Enable Javascript to view the ... PDF Open All Close All Description COL4A1 -related brain small-vessel disease is part of a group ...

  10. Distribution of cold adaptation proteins in microbial mats in Lake Joyce, Antarctica: Analysis of metagenomic data by using two bioinformatics tools.

    PubMed

    Koo, Hyunmin; Hakim, Joseph A; Fisher, Phillip R E; Grueneberg, Alexander; Andersen, Dale T; Bej, Asim K

    2016-01-01

    In this study, we report the distribution and abundance of cold-adaptation proteins in microbial mat communities in the perennially ice-covered Lake Joyce, located in the McMurdo Dry Valleys, Antarctica. We have used MG-RAST and R code bioinformatics tools on Illumina HiSeq2000 shotgun metagenomic data and compared the filtering efficacy of these two methods on cold-adaptation proteins. Overall, the abundance of cold-shock DEAD-box protein A (CSDA), antifreeze proteins (AFPs), fatty acid desaturase (FAD), trehalose synthase (TS), and cold-shock family of proteins (CSPs) were present in all mat samples at high, moderate, or low levels, whereas the ice nucleation protein (INP) was present only in the ice and bulbous mat samples at insignificant levels. Considering the near homogeneous temperature profile of Lake Joyce (0.08-0.29 °C), the distribution and abundance of these proteins across various mat samples predictively correlated with known functional attributes necessary for microbial communities to thrive in this ecosystem. The comparison of the MG-RAST and the R code methods showed dissimilar occurrences of the cold-adaptation protein sequences, though with insignificant ANOSIM (R = 0.357; p-value = 0.012), ADONIS (R(2) = 0.274; p-value = 0.03) and STAMP (p-values = 0.521-0.984) statistical analyses. Furthermore, filtering targeted sequences using the R code accounted for taxonomic groups by avoiding sequence redundancies, whereas the MG-RAST provided total counts resulting in a higher sequence output. The results from this study revealed for the first time the distribution of cold-adaptation proteins in six different types of microbial mats in Lake Joyce, while suggesting a simpler and more manageable user-defined method of R code, as compared to a web-based MG-RAST pipeline. PMID:26578243

  11. Medicine in the age of " Ulysses ": James Joyce's portrait of life, medicine, and disease on a Dublin day a century ago.

    PubMed

    Shanahan, Fergus; Quigley, Eamonn M

    2006-01-01

    Over time, contemporary writing becomes part of the historical record. In medicine, it is an important learning tool, particularly for understanding the experience and context of disease and illness. Although a century has elapsed since the fictional events on a single day described in James Joyce's Ulysses, the work is still fresh with references and allusions to doctors, illnesses, and the human experience. Ulysses provides perspective on medical and social history and offers a biting commentary of continuing relevance to the doctor-patient relationship. PMID:16702710

  12. Mutations in COL1A1 Gene Change Dentin Nanostructure.

    PubMed

    Duan, Xiaohong; Liu, Zhenxia; Gan, Yunna; Xia, Dan; Li, Qiang; Li, Yanling; Yang, Jiaji; Gao, Shan; Dong, Mingdong

    2016-04-01

    Although many studies have attempted to associate specific gene mutations with dentin phenotypic severity, it remains unknown how the mutations in COL1A1 gene influence the mechanical behavior of dentin collagen and matrix. Here, we reported one osteogenesis imperfecta (OI) pedigree caused by two new inserting mutations in exon 5 of COL1A1 (NM_000088.3:c.440_441insT;c.441_442insA), which resulted in the unstable expression of COL1A1 mRNA and half quantity of procollagen production. We investigated the morphological and mechanical features of proband's dentin using atomic force microscope (AFM), scanning electron microscope, and transmission electron microscope. Increased D-periodic spacing, variably enlarged collagen fibrils coating with fewer minerals were found in the mutated collagen. AFM analysis demonstrated rougher dentin surface and sparsely decreased Young's modulus in proband's dentin. We believe that our findings provide new insights into the genetic-/nano- mechanisms of dentin diseases, and may well explain OI dentin features with reduced mechanical strength and a lower crosslinked density. Anat Rec, 299:511-519, 2016. © 2015 Wiley Periodicals, Inc. PMID:26694865

  13. Dietary Lecithin Decreases Skeletal Muscle COL1A1 and COL3A1 Gene Expression in Finisher Gilts.

    PubMed

    Akit, Henny; Collins, Cherie; Fahri, Fahri; Hung, Alex; D'Souza, Daryl; Leury, Brian; Dunshea, Frank

    2016-01-01

    The purpose of this study was to investigate the effect of dietary lecithin on skeletal muscle gene expression of collagen precursors and enzymes involved in collagen synthesis and degradation. Finisher gilts with an average start weight of 55.9 ± 2.22 kg were fed diets containing either 0, 4, 20 or 80 g/kg soybean lecithin prior to harvest for six weeks and the rectus abdominis muscle gene expression profile was analyzed by quantitative real-time PCR. Lecithin treatment down-regulated Type I (α1) procollagen (COL1A1) and Type III (α1) procollagen (COL3A1) mRNA expression ( p < 0.05, respectively), indicating a decrease in the precursors for collagen synthesis. The α-subunit of prolyl 4-hydroxylase (P4H) mRNA expression also tended to be down-regulated ( p = 0.056), indicating a decrease in collagen synthesis. Decreased matrix metalloproteinase-1 (MMP-1) mRNA expression may reflect a positive regulatory response to the reduced collagen synthesis in muscle from the pigs fed lecithin ( p = 0.035). Lecithin had no effect on tissue inhibitor metalloproteinase-1 (TIMP-1), matrix metalloproteinase-13 (MMP-13) and lysyl oxidase mRNA expression. In conclusion, lecithin down-regulated COL1A1 and COL3A1 as well as tended to down-regulate α-subunit P4H expression. However, determination of muscle collagen content and solubility are required to support the gene functions. PMID:27338483

  14. Mutation survey and genotype-phenotype analysis of COL2A1 and COL11A1 genes in 16 Chinese patients with Stickler syndrome

    PubMed Central

    Wang, Xun; Jia, Xiaoyun; Xiao, Xueshan; Li, Shiqiang; Li, Jie; Li, Yadi; Wei, Yantao; Liang, Xiaoling

    2016-01-01

    Purpose To identify mutations in COL2A1 and COL11A1 genes and to examine the genotype-phenotype correlation in a cohort of Chinese patients with Stickler syndrome. Methods A total of 16 Chinese probands with Stickler syndrome were recruited, including nine with a family history of an autosomal dominant pattern and seven sporadic cases. All patients underwent full ocular and systemic examinations. Sanger sequencing was used to analyze all coding and adjacent regions of the COL2A1 and COL11A1 genes. Multiplex ligation-dependent probe amplification was performed to detect the gross indels of COL2A1 and COL11A1. Bioinformatics analysis was performed to evaluate the pathogenicity of the variants. Results Five mutations in COL2A1 were identified in six of 16 probands, including three novel (c.85C>T, c.3356delG, c.3401delG) mutations and two known mutations (c.1693C>T, c.2710C>T). Of the five mutations, three were truncated mutations, and the other two were missense mutations. Putative pathogenic mutations of the COL11A1 gene were absent in this cohort of patients. Gross indels were not found in COL2A1 or COL11A1 in any of the probands. High myopia was the most frequent initial ocular phenotype of Stickler syndrome. In this study, 12 Chinese probands lacked obvious systemic phenotypes. Conclusions In this study, three novel and two known mutations in the COL2A1 gene were identified in six of 16 Chinese patients with Stickler syndrome. This is the first study in a cohort of Chinese patients with Stickler syndrome, and the results expand the mutation spectrum of the COL2A1 gene. Analysis of the genotype-phenotype correlation showed that the early onset of high myopia with vitreous abnormalities may serve as a key indicator of Stickler syndrome, while the existence of mandibular protrusion in pediatric patients may be an efficient indicator for the absence of mutations in COL2A1 and COL11A1. PMID:27390512

  15. Col4a1 mutations cause progressive retinal neovascular defects and retinopathy

    PubMed Central

    Alavi, Marcel V.; Mao, Mao; Pawlikowski, Bradley T.; Kvezereli, Manana; Duncan, Jacque L.; Libby, Richard T.; John, Simon W. M.; Gould, Douglas B.

    2016-01-01

    Mutations in collagen, type IV, alpha 1 (COL4A1), a major component of basement membranes, cause multisystem disorders in humans and mice. In the eye, these include anterior segment dysgenesis, optic nerve hypoplasia and retinal vascular tortuosity. Here we investigate the retinal pathology in mice carrying dominant-negative Col4a1 mutations. To this end, we examined retinas longitudinally in vivo using fluorescein angiography, funduscopy and optical coherence tomography. We assessed retinal function by electroretinography and studied the retinal ultrastructural pathology. Retinal examinations revealed serous chorioretinopathy, retinal hemorrhages, fibrosis or signs of pathogenic angiogenesis with chorioretinal anastomosis in up to approximately 90% of Col4a1 mutant eyes depending on age and the specific mutation. To identify the cell-type responsible for pathogenesis we generated a conditional Col4a1 mutation and determined that primary vascular defects underlie Col4a1-associated retinopathy. We also found focal activation of Müller cells and increased expression of pro-angiogenic factors in retinas from Col4a1+/Δex41mice. Together, our findings suggest that patients with COL4A1 and COL4A2 mutations may be at elevated risk of retinal hemorrhages and that retinal examinations may be useful for identifying patients with COL4A1 and COL4A2 mutations who are also at elevated risk of hemorrhagic strokes. PMID:26813606

  16. ColE1 plasmid incompatibility: localization and analysis of mutations affecting incompatibility.

    PubMed Central

    Hashimoto-Gotoh, T; Inselburg, J

    1979-01-01

    Deletion mutants of plasmid ColE1 that involve the replication origin and adjacent regions of the plasmid have been studied to determine the mechanism by which those mutations affect the expression of plasmid incompatibility. It was observed that (i) a region of ColE1 that is involved in the expression of plasmid incompatibility lies between base pairs -185 and -684; (ii) the integrity of at least part of the region of ColE1 DNA between base pairs -185 and -572 is essential for the expression of ColE1 incompatibility; (iii) the expression of incompatibility is independent of the ability of the ColE1 genome to replicate autonomously; (iv) plasmid incompatibility is affected by plasmid copy number; and (v) ColE1 plasmid-mediated DNA replication of the lambda phage-ColE1 chimera lambda imm434 Oam29 Pam3 ColE1 is inhibited by ColE1-incompatible but not by ColE1-compatible plasmids. Images PMID:378980

  17. The transcription factor Lc-Maf participates in Col27a1 regulation during chondrocyte maturation

    SciTech Connect

    Mayo, Jaime L.; Holden, Devin N.; Barrow, Jeffery R.; Bridgewater, Laura C.

    2009-08-01

    The transcription factor Lc-Maf, which is a splice variant of c-Maf, is expressed in cartilage undergoing endochondral ossification and participates in the regulation of type II collagen through a cartilage-specific Col2a1 enhancer element. Type XXVII and type XI collagens are also expressed in cartilage during endochondral ossification, and so enhancer/reporter assays were used to determine whether Lc-Maf could regulate cartilage-specific enhancers from the Col27a1 and Col11a2 genes. The Col27a1 enhancer was upregulated over 4-fold by Lc-Maf, while the Col11a2 enhancer was downregulated slightly. To confirm the results of these reporter assays, rat chondrosarcoma (RCS) cells were transiently transfected with an Lc-Maf expression plasmid, and quantitative RT-PCR was performed to measure the expression of endogenous Col27a1 and Col11a2 genes. Endogenous Col27a1 was upregulated 6-fold by Lc-Maf overexpression, while endogenous Col11a2 was unchanged. Finally, in situ hybridization and immunohistochemistry were performed in the radius and ulna of embryonic day 17 mouse forelimbs undergoing endochondral ossification. Results demonstrated that Lc-Maf and Col27a1 mRNAs are coexpressed in proliferating and prehypertrophic regions, as would be predicted if Lc-Maf regulates Col27a1 expression. Type XXVII collagen protein was also most abundant in prehypertrophic and proliferating chondrocytes. Others have shown that mice that are null for Lc-Maf and c-Maf have expanded hypertrophic regions with reduced ossification and delayed vascularization. Separate studies have indicated that Col27a1 may serve as a scaffold for ossification and vascularization. The work presented here suggests that Lc-Maf may affect the process of endochondral ossification by participating in the regulation of Col27a1 expression.

  18. Strategy for prenatal diagnosis of osteogenesis imperfecta by linkage analysis to the type I collagen loci COL1A1 and COL1A2.

    PubMed

    Benušienė, E; Kučinskas, V

    2000-01-01

    To improve prenatal diagnosis of osteogenesis imperfecta (OI) in Lithuania, possibilities of indirect molecular genetic diagnosis were investigated in 11 families with dominant OI. Segregation of polymorphic DNA markers closely linked to COL1A1 and COL1A2 genes with OI phenotype was investigated. Polymorphic DNA markers applied were individual haplotypes constructed using a set of restriction enzyme sites within or close to the genes. Comparison of phenotypic features with the concordant collagen locus showed that in four pedigrees with OI Sillence type I segregated with COL1A1, while two pedigrees with OI Sillence type I and OI type IV segregated with COL1A2. Out of six remaining pedigrees with OI Sillence type I, three were concordant at both loci, two pedigrees were discordant at the locus COL1A2 and non-informative at the locus COL1A1 and one pedigree was concordant at the locus COL1A1 and non-informative at the locus COL1A2. Informativity of DNA markers applied was also investigated in the Lithuanian OI families. The frequencies of six restriction enzyme site dimorphisms in type I collagen loci were estimated and polymorphism information content (PIC) values were calculated for each restriction site and for a combination of three sites. COL1A1 locus dimorphisms A/MspI, B/RsaI and F/MnlI, showed PIC values of 0.327, 0.191 and 0.366, respectively, giving a combined PIC of 0.656 at the locus, while COL1A2 locus dimorphisms C/EcoRI, D/MspI and E/RsaI RFLPs had PIC values of 0.357, 0.168 and 0.331, respectively, giving a combined PIC of 0.655 at the locus. PMID:11208313

  19. RXTE monitoring of the intermediate polar TX Col

    NASA Astrophysics Data System (ADS)

    Wheatley, Peter J.

    I present a preliminary analysis of ongoing monthly X-ray observations of TX Col with RXTE. The lightcurves confirm that the relative strength of spin and beat modulations is highly variable in this system, and show that the timescale of variation is shorter than one month. Changes in these modulations are thought to represent a change in accretion geometry, possibly governed by a varying accretion rate. I find some evidence that the ratio of spin to beat amplitudes is correlated with the mean count rate.

  20. Dysspondyloenchondromatosis: Another COL2A1-Related Skeletal Dysplasia?

    PubMed Central

    Nakane, T.; Tando, T.; Aoyagi, K.; Hatakeyama, K.; Nishimura, G.; Coucke, I.P.J.; Mortier, G.; Sugita, K.

    2011-01-01

    Dysspondyloenchondromatosis (DSC) is a rare skeletal dysplasia that has currently been classified into the group of spondylometaphyseal dysplasias. To date, only 12 affected individuals have been reported. All cases are sporadic, and the etiology remains unknown. Distinctive features of DSC are anisospondyly and enchondroma-like lesions in the metaphyseal and diaphyseal portions of the long tubular bones. Affected individuals usually develop kyphoscoliosis and asymmetric limb shortening at an early age. Interestingly, some of the skeletal changes overlap with spondyloepimetaphyseal dysplasia (SEMD) Strudwick type, a rare type II collagen disorder. Based on this resemblance we postulated that DSC may be allelic to SEMD Strudwick type and therefore performed a COL2A1 analysis in an affected boy who was diagnosed as having DSC at the age of 3 years. The identification of a novel heterozygous COL2A1 missense mutation (p.Gly753Asp) in the proband confirms our hypothesis and suggests that DSC may be another type II collagen disorder. PMID:22570642

  1. In search of lost opportunities: Marcel Proyce and James Joust discuss doctors, diseases, life and death (a hypothetical conversation between Marcel Proust and James Joyce).

    PubMed

    Shanahan, Fergus; Quigley, Eamonn M

    2012-01-01

    Two of the most influential thinkers of the last century, Marcel Proust (1871-1922) and James Joyce (1882-1941), met in 1922 but had little to say to each other. Had circumstances been different, what might they have said? They had much in common, including an interest in doctors and diseases, but from different perspectives. Although the meeting was a lost opportunity to explore the insights of two great analysts of the human condition, their lives and works provide a record rich in detail and timeless in relevance. From this, we construct what might have been overheard during a conversation between Marcel Proyce and James Joust almost a century ago. PMID:22643723

  2. A Novel DHPLC-Based Procedure for the Analysis of COL1A1 and COL1A2 Mutations in Osteogenesis Imperfecta

    PubMed Central

    Fuccio, Antonella; Iorio, Mariangela; Amato, Felice; Elce, Ausilia; Ingino, Rosaria; Filocamo, Mirella; Castaldo, Giuseppe; Salvatore, Francesco; Tomaiuolo, Rossella

    2011-01-01

    Approximately 90% of patients with osteogenesis imperfecta (OI) exhibit dominant COL1A1 or COL1A2 mutations; however, molecular analysis is difficult because these genes span 51 and 52 exons, respectively. We devised a PCR-denaturing high-performance liquid chromatography (DHPLC) procedure to analyze the COL1A1 or COL1A2 coding regions and validated it using 130 DNA samples from individuals without OI, 25 DNA samples from two cells to investigate the procedure's potential for preimplantation diagnosis, and DNA samples from 10 patients with OI. Three novel intronic variants in vitro were expressed using a minigene assay to assess their effects on splicing. The procedure is rapid, inexpensive, and reproducible. Analysis of samples from individuals without OI revealed six novel and some known polymorphisms useful for linkage diagnosis because of high heterozygosity. Analysis of two-cell samples confirmed the known genotype in 24 of 25 experiments; DNA failed to amplify in only one case. No incidence of allele dropout was recorded. DHPLC revealed six novel mutations, three of which were intronic, in all patients with OI, and these results were confirmed by means of COL1A1 and COL1A2 direct sequencing. Expression of intronic mutations demonstrated that variant 804 + 2_804 + 3delTG in intron 11 disrupts normal splicing, thereby leading to formation of two alternative products. Variants c.3046-4_3046-5dupCT (COL1A1) and c.891 + 77A>T (COL1A2) did not affect splicing. The described DHPLC protocol combined with the minigene assay may contribute to molecular diagnosis in OI. Moreover, this protocol will aid in counseling about prenatal and preimplantation diagnosis. PMID:21884818

  3. Differentiation of Col I and Col III Isoforms in Stromal Models of Ovarian Cancer by Analysis of Second Harmonic Generation Polarization and Emission Directionality

    PubMed Central

    Tilbury, Karissa; Lien, Chi-Hsiang; Chen, Shean-Jen; Campagnola, Paul J.

    2014-01-01

    A profound remodeling of the extracellular matrix occurs in many epithelial cancers. In ovarian cancer, the minor collagen isoform of Col III becomes upregulated in invasive disease. Here we use second harmonic generation (SHG) imaging microscopy to probe structural differences in fibrillar models of the ovarian stroma comprised of mixtures of Col I and III. The SHG intensity and forward-backward ratios decrease with increasing Col III content, consistent with decreased phasematching due to more randomized structures. We further probe the net collagen α-helix pitch angle within the gel mixtures using what is believed to be a new pixel-based polarization-resolved approach that combines and extends previous analyses. The extracted pitch angles are consistent with those of peptide models and the method has sufficient sensitivity to differentiate Col I from the Col I/Col III mixtures. We further developed the pixel-based approach to extract the SHG signal polarization anisotropy from the same polarization-resolved image matrix. Using this approach, we found that increased Col III results in decreased alignment of the dipole moments within the focal volume. Collectively, the SHG measurements and analysis all indicate that incorporation of Col III results in decreased organization across several levels of collagen organization. Furthermore, the findings suggest that the collagen isoforms comingle within the same fibrils, in good agreement with ultrastructural data. The pixel-based polarization analyses (both excitation and emission) afford determination of structural properties without the previous requirement of having well-aligned fibers, and the approaches should be generally applicable in tissue. PMID:24461010

  4. Downregulation of let‑7b promotes COL1A1 and COL1A2 expression in dermis and skin fibroblasts during heat wound repair.

    PubMed

    Liu, Jinyan; Luo, Chengqun; Yin, Zhaoqi; Li, Ping; Wang, Shaohua; Chen, Jia; He, Quanyong; Zhou, Jianda

    2016-03-01

    MicroRNAs (miRs), a class of non‑coding RNAs 18‑25 nucleotides in length, generally serve suppressive role in the regulation of gene expression via directly binding to the 3'‑untranslated region (UTR) of their target mRNA. Previous studies have identified several miRs to be involved in thermal injury repair. However, the role of miR let‑7b during the recovery of thermal injury, in addition to the underlying mechanisms, has not previously been studied. In the present study, the expression of let‑7b was observed to be significantly increased in skin tissue shortly following thermal injury, however, gradually reduced during the recovery of thermal injury. Notably, similar findings were observed in heat‑denatured skin fibroblasts. Furthermore, collagen, type I, alpha 1 (COL1A1) and collagen, type I, alpha 2 (COL1A2), which are associated with the synthesis of type I collagen, were identified as two targets of let‑7b in skin fibroblasts. The overexpression of let‑7b was observed to upregulate the protein expression levels of COL1A1 and COL1A2, while knockdown of let‑7b reduced the levels of COL1A1 and COL1A2 in skin fibroblasts. Furthermore, COL1A1 and COL1A2 were significantly downregulated shortly following thermal injury, while gradually upregulated during healing, in heat‑damaged skin tissue and skin fibroblasts, with the expression profiles opposite to that of let‑7b. Taken together, this suggests that the downregulation of let‑7b in heat‑damaged dermis promotes the synthesis of type I collagen and thus aids in burn wound repair. PMID:26861712

  5. Differentiation of Col I and Col III isoforms in stromal models of ovarian cancer by analysis of second harmonic generation polarization and emission directionality.

    PubMed

    Tilbury, Karissa; Lien, Chi-Hsiang; Chen, Shean-Jen; Campagnola, Paul J

    2014-01-21

    A profound remodeling of the extracellular matrix occurs in many epithelial cancers. In ovarian cancer, the minor collagen isoform of Col III becomes upregulated in invasive disease. Here we use second harmonic generation (SHG) imaging microscopy to probe structural differences in fibrillar models of the ovarian stroma comprised of mixtures of Col I and III. The SHG intensity and forward-backward ratios decrease with increasing Col III content, consistent with decreased phasematching due to more randomized structures. We further probe the net collagen α-helix pitch angle within the gel mixtures using what is believed to be a new pixel-based polarization-resolved approach that combines and extends previous analyses. The extracted pitch angles are consistent with those of peptide models and the method has sufficient sensitivity to differentiate Col I from the Col I/Col III mixtures. We further developed the pixel-based approach to extract the SHG signal polarization anisotropy from the same polarization-resolved image matrix. Using this approach, we found that increased Col III results in decreased alignment of the dipole moments within the focal volume. Collectively, the SHG measurements and analysis all indicate that incorporation of Col III results in decreased organization across several levels of collagen organization. Furthermore, the findings suggest that the collagen isoforms comingle within the same fibrils, in good agreement with ultrastructural data. The pixel-based polarization analyses (both excitation and emission) afford determination of structural properties without the previous requirement of having well-aligned fibers, and the approaches should be generally applicable in tissue. PMID:24461010

  6. Expression of COL6A1 predicts prognosis in cervical cancer patients

    PubMed Central

    Hou, Teng; Tong, Chongjie; Kazobinka, Gallina; Zhang, Weijing; Huang, Xin; Huang, Yongwen; Zhang, Yanna

    2016-01-01

    COL6A1 has been shown to play an important role in tumor initiation and progression. The present study is to investigate the clinical significance of COL6A1 in cervical cancer. In this study, the COL6A1 expression levels in 10 paired cervical cancer tissues and the adjacent non-tumor tissues were examined by real-time PCR. The expression of COL6A1 protein was examined in 162 cervical cancer samples by immunohistochemistry, and the correlation of COL6A1 expression with clinicopathologic factors was analyzed. The overall and recurrent-free survival rates were estimated using Kaplan-Meier method and compared with the log-rank test. The prognostic analysis was carried out with multivariate Cox regressions model. The result showed that COL6A1 expression was up-regulated in cervical cancer tissues in compared with that in non-tumor tissues. High expression of COL6A1 was significantly correlated with FIGO stage (P<0.001), tumor size (P=0.025) and lymph node metastasis (P=0.028) of the disease. Moreover, survival analysis showed that high expression of COL6A1 was significantly associated with poorer overall (OS) and recurrent free (RFS) survival (p=0.004 and =0.001, respectively) of cervical cancer patients. Multivariate analysis suggested that COL6A1 expression was an independent prognostic marker of cervical cancer (P=0.029). Thus, COL6A1 may serve as an oncogene in the initiation and progression of cervical cancer, and as a predictor of poor prognosis in cervical cancer patients. PMID:27398167

  7. Clinical and molecular characterization of 40 patients with classic Ehlers–Danlos syndrome: identification of 18 COL5A1 and 2 COL5A2 novel mutations

    PubMed Central

    2013-01-01

    Background Classic Ehlers–Danlos syndrome (cEDS) is a rare autosomal dominant connective tissue disorder that is primarily characterized by skin hyperextensibility, abnormal wound healing/atrophic scars, and joint hypermobility. A recent study demonstrated that more than 90% of patients who satisfy all of these major criteria harbor a type V collagen (COLLV) defect. Methods This cohort included 40 patients with cEDS who were clinically diagnosed according to the Villefranche nosology. The flowchart that was adopted for mutation detection consisted of sequencing the COL5A1 gene and, if no mutation was detected, COL5A2 analysis. In the negative patients the presence of large genomic rearrangements in COL5A1 was investigated using MLPA, and positive results were confirmed via SNP-array analysis. Results We report the clinical and molecular characterization of 40 patients from 28 families, consisting of 14 pediatric patients and 26 adults. A family history of cEDS was present in 9 patients. The majority of the patients fulfilled all the major diagnostic criteria for cEDS; atrophic scars were absent in 2 females, skin hyperextensibility was not detected in a male and joint hypermobility was negative in 8 patients (20% of the entire cohort). Wide inter- and intra-familial phenotypic heterogeneity was observed. We identified causal mutations with a detection rate of approximately 93%. In 25/28 probands, COL5A1 or COL5A2 mutations were detected. Twenty-one mutations were in the COL5A1 gene, 18 of which were novel (2 recurrent). Of these, 16 mutations led to nonsense-mediated mRNA decay (NMD) and to COLLV haploinsufficiency and 5 mutations were structural. Two novel COL5A2 splice mutations were detected in patients with the most severe phenotypes. The known p. (Arg312Cys) mutation in the COL1A1 gene was identified in one patient with vascular-like cEDS. Conclusions Our findings highlight that the three major criteria for cEDS are useful and sufficient for cEDS clinical

  8. Splicing analysis of unclassified variants in COL2A1 and COL11A1 identifies deep intronic pathogenic mutations

    PubMed Central

    Richards, Allan J; McNinch, Annie; Whittaker, Joanne; Treacy, Becky; Oakhill, Kim; Poulson, Arabella; Snead, Martin P

    2012-01-01

    UK NHS diagnostic service sequence analysis of genes generally examines and reports on variations within a designated region 5′ and 3′ of each exon, typically 30 bp up and downstream. However, because of the degenerate nature of the splice sites, intronic variants outside the AG and GT dinucleotides of the acceptor and donor splice sites (ASS and DSS) are most often classified as being of unknown clinical significance, unless there is some functional evidence of their pathogenicity. It is now becoming clear that mutations deep within introns can also interfere with normal processing of pre-mRNA and result in pathogenic effects on the mature transcript. In diagnostic laboratories, these deep intronic variants most often fall outside of the regions analysed and so are rarely reported. With the likelihood that next generation sequencing will identify more of these unclassified variants, it will become important to perform additional studies to determine the pathogenicity of such sequence anomalies. Here, we analyse variants detected in either COL2A1 or COL11A1 in patients with Stickler syndrome. These have been analysed both in silico and functionally using either RNA isolated from the patient's cells or, more commonly, minigenes as splicing reporters. We show that deep intronic mutations are not a rare occurrence, including one variant that results in multiple transcripts, where both de novo donor and ASS are created by the mutation. Another variant produces transcripts that result in either haploinsufficiency or a dominant negative effect, potentially modifying the disease phenotype. PMID:22189268

  9. Consistent linkage of dominantly inherited osteogenesis imperfecta to the type I collagen loci: COL1A1 and COL1A2.

    PubMed

    Sykes, B; Ogilvie, D; Wordsworth, P; Wallis, G; Mathew, C; Beighton, P; Nicholls, A; Pope, F M; Thompson, E; Tsipouras, P

    1990-02-01

    The segregation of COL1A1 and COL1A2, the two genes which encode the chains of type I collagen, was analyzed in 38 dominant osteogenesis imperfecta (OI) pedigrees by using polymorphic markers within or close to the genes. This was done in order to estimate the consistency of linkage of OI genes to these two loci. None of the 38 pedigrees showed evidence of recombination between the OI gene and both collagen loci, suggesting that the frequency of unlinked loci in the population must be low. From these results, approximate 95% confidence limits for the proportion of families linked to the type I collagen genes can be set between .91 and 1.00. This is high enough to base prenatal diagnosis of dominantly inherited OI on linkage to these genes even in families which are too small for the linkage to be independently confirmed to high levels of significance. When phenotypic features were compared with the concordant collagen locus, all eight pedigrees with Sillence OI type IV segregated with COL1A2. On the other hand, Sillence OI type I segregated with both COL1A1 (17 pedigrees) and COL1A2 (7 pedigrees). The concordant locus was uncertain in the remaining six OI type I pedigrees. Of several other features, the presence or absence of presenile hearing loss was the best predictor of the mutant locus in OI type I families, with 13 of the 17 COL1A1 segregants and none of the 7 COL1A2 segregants showing this feature. PMID:1967900

  10. Linkage approach and direct COL4A5 gene mutation screening in Alport syndrome

    SciTech Connect

    Turco, A.E.; Rossetti, S.; Biasi, O.

    1994-09-01

    Alport Syndrome (AS) is transmitted as an X-linked dominant trait in the majority of families, the defective gene being COL4A5 at Xq22. In the remaining cases AS appears to be autosomally inherited. Recently, mutations in COL4A3 and COL4A4 genes at 2q35-q37 were identified in families with autosomal recessive AS. Mutation detection screening is being performed by non-radioactive single stand conformation polymorphism (SSCP), heteroduplex analysis, and automated DNA sequencing in over 170 AS patients enrolled in the ongoing Italian Multicenter Study on AS. So far twenty-five different mutations have been found, including missense, splicing, and frameshifts. Moreover, by using six tightly linked COL4A5 informative makers, we have also typed two larger AS families, and have shown compatible sex-linked transmission in one other, suggesting autosomal recessive inheritance. In this latter three-generation COL4A5-unlinked family we are now looking for linkage and for mutations in the candidate COL4A3 and COL4A4 genes on chromosome 2q.

  11. Craniofacial and Dental Defects in the Col1a1Jrt/+ Mouse Model of Osteogenesis Imperfecta.

    PubMed

    Eimar, H; Tamimi, F; Retrouvey, J-M; Rauch, F; Aubin, J E; McKee, M D

    2016-07-01

    Certain mutations in the COL1A1 and COL1A2 genes produce clinical symptoms of both osteogenesis imperfecta (OI) and Ehlers-Danlos syndrome (EDS) that include abnormal craniofacial growth, dental malocclusion, and dentinogenesis imperfecta. A mouse model (Col1a1(Jrt)/+) was recently developed that had a skeletal phenotype and other features consistent with moderate-to-severe OI and also with EDS. The craniofacial phenotype of 4- and 20-wk-old Col1a1(Jrt)/+ mice and wild-type littermates was assessed by micro-computed tomography (µCT) and morphometry. Teeth and the periodontal ligament compartment were analyzed by µCT, light microscopy/histomorphometry, and electron microscopy. Over time, at 20 wk, Col1a1(Jrt)/+ mice developed smaller heads, a shortened anterior cranial base, class III occlusion, and a mandibular side shift with shorter morphology in the masticatory region (maxilla and mandible). Col1a1(Jrt)/+ mice also had changes in the periodontal compartment and abnormalities in the dentin matrix and mineralization. These findings validate Col1a1(Jrt)/+ mice as a model for OI and EDS in humans. PMID:26951553

  12. Characterizing Microbial Mat Morphology with Structure from Motion Techniques in Ice-Covered Lake Joyce, McMurdo Dry Valleys, Antarctica

    NASA Astrophysics Data System (ADS)

    Mackey, T. J.; Leidman, S. Z.; Allen, B.; Hawes, I.; Lawrence, J.; Jungblut, A. D.; Krusor, M.; Coleman, L.; Sumner, D. Y.

    2015-12-01

    Structure from Motion (SFM) techniques can provide quantitative morphological documentation of otherwise inaccessible benthic ecosystems such as microbial mats in Lake Joyce, a perennially ice-covered lake of the Antarctic McMurdo Dry Valleys (MDV). Microbial mats are a key ecosystem of MDV lakes, and diverse mat morphologies like pinnacles emerge from interactions among microbial behavior, mineralization, and environmental conditions. Environmental gradients can be isolated to test mat growth models, but assessment of mat morphology along these gradients is complicated by their inaccessibility: the Lake Joyce ice cover is 4-5 m thick, water depths containing diverse pinnacle morphologies are 9-14 m, and relevant mat features are cm-scale. In order to map mat pinnacle morphology in different sedimentary settings, we deployed drop cameras (SeaViewer and GoPro) through 29 GPS referenced drill holes clustered into six stations along a transect spanning 880 m. Once under the ice cover, a boom containing a second GoPro camera was unfurled and rotated to collect oblique images of the benthic mats within dm of the mat-water interface. This setup allowed imaging from all sides over a ~1.5 m diameter area of the lake bottom. Underwater lens parameters were determined for each camera in Agisoft Lens; images were reconstructed and oriented in space with the SFM software Agisoft Photoscan, using the drop camera axis of rotation as up. The reconstructions were compared to downward facing images to assess accuracy, and similar images of an object with known geometry provided a test for expected error in reconstructions. Downward facing images identify decreasing pinnacle abundance in higher sedimentation settings, and quantitative measurements of 3D reconstructions in KeckCAVES LidarViewer supplement these mat morphological facies with measurements of pinnacle height and orientation. Reconstructions also help isolate confounding variables for mat facies trends with measurements

  13. Collagen type IV-related nephropathies in Portugal: pathogenic COL4A5 mutations and clinical characterization of 22 families.

    PubMed

    Nabais Sá, M J; Sampaio, S; Oliveira, A; Alves, S; Moura, C P; Silva, S E; Castro, R; Araújo, J A; Rodrigues, M; Neves, F; Seabra, J; Soares, C; Gaspar, M A; Tavares, I; Freitas, L; Sousa, T C; Henriques, A C; Costa, F T; Morgado, E; Sousa, F T; Sousa, J P; da Costa, A G; Filipe, R; Garrido, J; Montalban, J; Ponce, P; Alves, R; Faria, B; Carvalho, M F; Pestana, M; Carvalho, F; Oliveira, J P

    2015-11-01

    Alport syndrome (AS) is caused by pathogenic mutations in the genes encoding α3, α4 or α5 chains of collagen IV (COL4A3/COL4A4/COL4A5), resulting in hematuria, chronic renal failure (CRF), sensorineural hearing loss (SNHL) and ocular abnormalities. Mutations in the X-linked COL4A5 gene have been identified in 85% of the families (XLAS). In this study, 22 of 60 probands (37%) of unrelated Portuguese families, with clinical diagnosis of AS and no evidence of autosomal inheritance, had pathogenic COL4A5 mutations detected by Sanger sequencing and/or multiplex-ligation probe amplification, of which 12 (57%) are novel. Males had more severe and earlier renal and extrarenal complications, but microscopic hematuria was a constant finding irrespective of gender. Nonsense and splice site mutations, as well as small and large deletions, were associated with younger age of onset of SNHL in males, and with higher risk of CRF and SNHL in females. Pathogenic COL4A3 or COL4A4 mutations were subsequently identified in more than half of the families without a pathogenic mutation in COL4A5. The lower than expected prevalence of XLAS in Portuguese families warrants the use of next-generation sequencing for simultaneous COL4A3/COL4A4/COL4A5 analysis, as first-tier approach to the genetic diagnosis of collagen type IV-related nephropathies. PMID:25307721

  14. Experimental murine myopia induces collagen type Iα1 (COL1A1) DNA methylation and altered COL1A1 messenger RNA expression in sclera

    PubMed Central

    Zhou, Xiangtian; Ji, Fengtao; An, Jianhong; Zhao, Fuxin; Shi, Fanjun; Huang, Furong; Li, Yuan; Jiao, Shiming; Yan, Dongsheng; Chen, Xiaoyan; Chen, JiangFan

    2012-01-01

    Purpose To investigate whether myopia development is associated with changes of scleral DNA methylation in cytosine-phosphate-guanine (CpG) sites in the collagen 1A1 (COL1A1) promoter and messenger RNA (mRNA) levels following murine form deprivation myopia. Methods Fifty-seven C57BL/6 mice (postnatal day 23) were randomly assigned to four groups: (1) monocular form deprivation (MD) in which a diffuser lens was placed over one eye for 28 days; (2) normal controls without MD; (3) MD recovery in which the diffuser lens was removed for seven days; and (4) MD recovery normal controls. The DNA methylation pattern in COL1A1 promoter and exon 1 was determined by bisulfite DNA sequencing, and the COL1A1 mRNA level in sclera was determined by quantitative PCR. Results MD was found to induce myopia in the treated eyes. Six CpG sites in the promoter and exon 1 region of COL1A1 were methylated with significantly higher frequency in the treated eyes than normal control eyes (p<0.05), with CpG island methylation in MD-contralateral eyes being intermediate. Consistent with the CpG methylation, scleral COL1A1 mRNA was reduced by 57% in the MD-treated eyes compared to normal controls (p<0.05). After seven days of MD recovery, CpG methylation was significantly reduced (p=0.01). The methylation patterns returned to near normal level in five CpG sites, but the sixth was hypomethylated compared to normal controls. Conclusions In parallel with the development of myopia and the reduced COL1A1 mRNA, the frequency of methylation in CpG sites of the COL1A1 promoter/exon 1 increased during MD and returned to near normal during recovery. Thus, hypermethylation of CpG sites in the promoter/exon 1 of COL1A1 may underlie reduced collagen synthesis at the transcriptional level in myopic scleras. PMID:22690110

  15. Novel mutations in the COL2A1 gene in Japanese patients with Stickler syndrome.

    PubMed

    Kondo, Hiroyuki; Matsushita, Itsuka; Nagata, Tatsuo; Hayashi, Takaaki; Kakinoki, Masashi; Uchio, Eiichi; Kondo, Mineo; Ohji, Masahito; Kusaka, Shunji

    2016-01-01

    Stickler syndrome is an inherited connective tissue disorder that affects the eyes, cartilage and articular tissues. The phenotypes of Stickler syndrome include congenital high myopia, retinal detachment, premature joint degeneration, hearing impairment and craniofacial anomalies, such as cleft palate and midline facial hypoplasia. The disease is genetically heterogeneous, and the majority of the cases are caused by mutations in the COL2A1 gene. We examined 40 Japanese patients with Stickler syndrome from 23 families to determine whether they had mutations in the COL2A1 gene. This analysis was conducted by examining each patient's genomic DNA by Sanger sequencing. Five nonsense, 4 splicing and 8 deletion mutations in the COL2A1 gene were identified, accounting for 21 of the 23 families. Different mutations of the COL2A1 gene were associated with similar phenotypes but with different degrees of expressivity. PMID:27408751

  16. Novel mutations in the COL2A1 gene in Japanese patients with Stickler syndrome

    PubMed Central

    Kondo, Hiroyuki; Matsushita, Itsuka; Nagata, Tatsuo; Hayashi, Takaaki; Kakinoki, Masashi; Uchio, Eiichi; Kondo, Mineo; Ohji, Masahito; Kusaka, Shunji

    2016-01-01

    Stickler syndrome is an inherited connective tissue disorder that affects the eyes, cartilage and articular tissues. The phenotypes of Stickler syndrome include congenital high myopia, retinal detachment, premature joint degeneration, hearing impairment and craniofacial anomalies, such as cleft palate and midline facial hypoplasia. The disease is genetically heterogeneous, and the majority of the cases are caused by mutations in the COL2A1 gene. We examined 40 Japanese patients with Stickler syndrome from 23 families to determine whether they had mutations in the COL2A1 gene. This analysis was conducted by examining each patient’s genomic DNA by Sanger sequencing. Five nonsense, 4 splicing and 8 deletion mutations in the COL2A1 gene were identified, accounting for 21 of the 23 families. Different mutations of the COL2A1 gene were associated with similar phenotypes but with different degrees of expressivity. PMID:27408751

  17. Persistent Notochord in a Fetus with COL2A1 Mutation

    PubMed Central

    Codsi, Elisabeth; Brost, Brian C.; Faksh, Arij; Volk, Amber K.; Borowski, Kristi S.

    2015-01-01

    Multiple anomalies including micromelia, poor mineralization of the vertebrae, and a persistent notochord were identified on second trimester ultrasound in a fetus with a COL2A1 mutation. To our knowledge, this represents the first case of a persistent notochord associated with a COL2A1 mutation in humans. In this case report, we describe ultrasound and postmortem findings and review the pathogenesis associated with a persistent notochord. PMID:26435866

  18. Is the COL5A1 rs12722 gene polymorphism associated with running economy?

    PubMed

    Bertuzzi, Rômulo; Pasqua, Leonardo A; Bueno, Salomão; Lima-Silva, Adriano Eduardo; Matsuda, Monique; Marquezini, Monica; Saldiva, Paulo H

    2014-01-01

    The COL5A1 rs12722 polymorphism is considered to be a novel genetic marker for endurance running performance. It has been postulated that COL5A1 rs12722 may influence the elasticity of tendons and the energetic cost of running. To date, there are no experimental data in the literature supporting the relationship between range of motion, running economy, and the COL5A1 rs12722 gene polymorphism. Therefore, the main purpose of the current study was to analyze the influence of the COL5A1rs12722 polymorphism on running economy and range of motion. One hundred and fifty (n = 150) physically active young men performed the following tests: a) a maximal incremental treadmill test, b) two constant-speed running tests (10 km · h(-1)) and 12 km · h(-1)) to determine the running economy, and c) a sit-and-reach test to determine the range of motion. All of the subjects were genotyped for the COL5A1 rs12722 single-nucleotide polymorphism. The genotype frequencies were TT = 27.9%, CT = 55.8%, and CC = 16.3%. There were no significant differences between COL5A1 genotypes for running economy measured at 10 km · h(-1) (p = 0.232) and 12 km · h(-1) (p = 0.259). Similarly, there were no significant differences between COL5A1 genotypes for range of motion (p = 0.337). These findings suggest that the previous relationship reported between COL5A1 rs12722 genotypes and running endurance performance might not be mediated by the energetic cost of running. PMID:25188268

  19. Is the COL5A1 rs12722 Gene Polymorphism Associated with Running Economy?

    PubMed Central

    Bertuzzi, Rômulo; Pasqua, Leonardo A.; Bueno, Salomão; Lima-Silva, Adriano Eduardo; Matsuda, Monique; Marquezini, Monica; Saldiva, Paulo H.

    2014-01-01

    The COL5A1 rs12722 polymorphism is considered to be a novel genetic marker for endurance running performance. It has been postulated that COL5A1 rs12722 may influence the elasticity of tendons and the energetic cost of running. To date, there are no experimental data in the literature supporting the relationship between range of motion, running economy, and the COL5A1 rs12722 gene polymorphism. Therefore, the main purpose of the current study was to analyze the influence of the COL5A1rs12722 polymorphism on running economy and range of motion. One hundred and fifty (n = 150) physically active young men performed the following tests: a) a maximal incremental treadmill test, b) two constant-speed running tests (10 km•h−1 and 12 km•h−1) to determine the running economy, and c) a sit-and-reach test to determine the range of motion. All of the subjects were genotyped for the COL5A1 rs12722 single-nucleotide polymorphism. The genotype frequencies were TT = 27.9%, CT = 55.8%, and CC = 16.3%. There were no significant differences between COL5A1 genotypes for running economy measured at 10 km•h−1 (p = 0.232) and 12 km•h−1 (p = 0.259). Similarly, there were no significant differences between COL5A1 genotypes for range of motion (p = 0.337). These findings suggest that the previous relationship reported between COL5A1 rs12722 genotypes and running endurance performance might not be mediated by the energetic cost of running. PMID:25188268

  20. Association of COL4A1 gene polymorphisms with cerebral palsy in a Chinese Han population.

    PubMed

    Bi, D; Wang, H; Shang, Q; Xu, Y; Wang, F; Chen, M; Ma, C; Sun, Y; Zhao, X; Gao, C; Wang, L; Zhu, C; Xing, Q

    2016-08-01

    The basement membrane (BM) is an extracellular matrix associated with overlying cells and is important for proper tissue development, stability, and physiology. COL4A1 is the most abundant component of type IV collagen in the BM, and COL4A1 variants can present with variable phenotypes that might be related to cerebral palsy (CP). We postulated, therefore, that variations in the COL4A1 gene might play an important role in the etiology of CP. In this study, six single nucleotide polymorphisms (SNPs) in the COL4A1 gene were genotyped among 351 CP patients and 220 healthy controls from the Chinese Han population. Significant association was found for an association between CP and rs1961495 (allele: p = 0.008, odds ratio (OR) = 1.387, 95% confidence interval (CI) = 1.088-1.767) and rs1411040 (allele: p = 0.009, OR = 1.746, 95% CI = 1.148-2.656) SNPs of the COL4A1 gene. Multifactor dimensionality reduction analysis suggested that these SNPs had interactive effects on the risk of CP. This study is the first attempt to investigate the contribution of polymorphisms in the COL4A1 gene to the susceptibility of CP in a Chinese Han population. This study shows an association of the COL4A1 gene with CP and suggests a potential role of COL4A1 in the pathogenesis of CP. PMID:26748532

  1. The COL5A1 genotype is associated with range of motion

    PubMed Central

    Lim, Seung-Taek; Kim, Chang-Sun; Kim, Woo-Nam; Min, Seok-Ki

    2015-01-01

    [Purpose] The aim of our study was to investigate the association between COL5A1 genotype and Range of Motion as measured by the passive straight leg raise (SLR) and whole body join laxity (WBJL) in Asian population. [Methods] One hundred and seventy seven participants including Korean and Japanese college students (male = 109, female = 68) participated in the study. Each subject performed the passive straight leg raise and whole body join laxity test. Genotyping for the COL5A1 (rs 12722) polymorphism was performed using the TaqMan approach. The COL5A1 genotype exhibited a Hardy-Weinberg equilibrium distribution in our population. [Results] The physical parameters including height, weight, and BMI were higher in < 90° group than > 90° group. The SLR exhibited significant difference among the COL5A1 group. However, the WBJL did not differ significantly among the COL5A1 genotype, but significant difference was seen in CC genotype when compared to CT (2.99 ± 1.72) or TT (2.70 ± 1.52) genotype. [Conclusion] We concluded that COL5A1 gene polymorphism is associated with increased SLR ROM in Asian population. PMID:26244122

  2. Association of COL2A1 Gene Polymorphism with Degenerative Lumbar Scoliosis

    PubMed Central

    Hwang, Dae Woo; Lee, Sang Hoon; Kim, Jung Youn; Kim, Dong Hwan

    2014-01-01

    Background Degenerative lumbar scoliosis (DLS) progresses with aging after 50-60 years, and the genetic association of DLS remains largely unclear. In this study, the genetic association between collagen type II alpha 1 (COL2A1) gene and DLS was investigated. Methods COL2A1 gene polymorphism was investigated in DLS subjects compared to healthy controls to investigate the possibility of its association with COL2A1 gene. Based on a single nucleotide polymorphism (SNP) database, SNP (rs2276454) in COL2A1 were selected and genotyped using direct sequencing in 51 patients with DLS and 235 healthy controls. The SNP effects were analyzed using three models of codominant, dominant, and recessive. Logistic regression models were calculated for odds ratios (ORs) with 95% confidence intervals (CIs) and corresponding p-values, controlling age and gender as co-variables. Results SNP (rs2276454) in COL2A1 was significantly associated with the degenerative lumbar scoliosis in the codominant (OR, 1.90; 95% CI, 1.17 to 3.10; p = 0.008) and dominant models (OR, 3.58; 95% CI, 1.59 to 9.29; p = 0.001). Conclusions The results suggest that COL2A1 is associated with the risk of DLS in Korean population. PMID:25436060

  3. Allele dependent silencing of COL1A2 using small interfering RNAs

    PubMed Central

    Lindahl, Katarina; Rubin, Carl-Johan; Kindmark, Andreas; Ljunggren, Östen

    2008-01-01

    Osteogenesis imperfecta (OI) is generally caused by a dominant mutation in Collagen I, encoded by the genes COL1A1 and COL1A2. To date there is no satisfactory therapy for OI, but inactivation of the mutant allele through small interfering RNAs (siRNA) is a promising approach, as siRNAs targeting each allele of a polymorphism could be used for allele-specific silencing irrespective of the location of the actual mutations. In this study we examined the allele dependent effects of several tiled siRNAs targeting a region surrounding an exonic COL1A2 T/C polymorphism (rs1800222) in heterozygous primary human bone cells. Relative abundances of COL1A2 alleles were determined by cDNA sequencing and overall COL1A2 abundance was analyzed by quantitative PCR. One of the siRNAs decreased overall COL1A2 abundance by 71% of which 75% was due to silencing of the targeted T-allele. In conclusion, allele-preferential silencing of Collagen type I genes may be a future therapeutic approach for OI. PMID:19015742

  4. Reactive stroma component COL6A1 is upregulated in castration-resistant prostate cancer and promotes tumor growth

    PubMed Central

    Shen, Yi-Jun; Wang, Hong-Kai; Zhang, Gui-Ming; Ye, Ding-Wei

    2015-01-01

    Castration-resistant prostate cancer (CRPC) remains the most critical challenge in the clinical management of prostate cancer (PCa). Reactive stromal changes in PCa are likely involved in the emergence of CRPC. In the present study, we identified a novel oncogene termed COL6A1 which was upregulated in the reactive stroma of CRPC. We established an androgen-independent LNCaP (LNCaP-AI) cell line in steroid-reduced (SR) medium within 2 months. We examined COL6A1 expression with western blot during the LNCaP-AI induction, and studied the function of COL6A1 in vitro and in vivo. Immunohistochemical staining of COL6A1 was performed in ten pairs of androgen-sensitive PCa and CRPC samples. We demonstrated that COL6A1 expression was markedly increased in LNCaP-AI cells and CRPC tissues compared with LNCaP cells and paired androgen-sensitive PCa specimens. In vitro, COL6A1 knockdown resulted in G1-S cell cycle arrest and descended vitality. Overexpression of COL6A1 was associated with accelerated S phase entry and elevated vitality in prostate cancer cells. COL6A1 also promoted tumorigenesis of LNCaP cells in vivo. Taken together, these data suggest an important role of COL6A1 in the molecular etiology of castration-resistant prostate cancer, and support the potential use of COL6A1 in CRPC therapy. PMID:25895032

  5. Identification and characterization of the novel Col10a1 regulatory mechanism during chondrocyte hypertrophic differentiation.

    PubMed

    Gu, J; Lu, Y; Li, F; Qiao, L; Wang, Q; Li, N; Borgia, J A; Deng, Y; Lei, G; Zheng, Q

    2014-01-01

    The majority of human skeleton develops through the endochondral pathway, in which cartilage-forming chondrocytes proliferate and enlarge into hypertrophic chondrocytes that eventually undergo apoptosis and are replaced by bone. Although at a terminal differentiation stage, hypertrophic chondrocytes have been implicated as the principal engine of bone growth. Abnormal chondrocyte hypertrophy has been seen in many skeletal dysplasia and osteoarthritis. Meanwhile, as a specific marker of hypertrophic chondrocytes, the type X collagen gene (COL10A1) is also critical for endochondral bone formation, as mutation and altered COL10A1 expression are often accompanied by abnormal chondrocyte hypertrophy in many skeletal diseases. However, how the type X collagen gene is regulated during chondrocyte hypertrophy has not been fully elucidated. We have recently demonstrated that Runx2 interaction with a 150-bp mouse Col10a1 cis-enhancer is required but not sufficient for its hypertrophic chondrocyte-specific reporter expression in transgenic mice, suggesting requirement of additional Col10a1 regulators. In this study, we report in silico sequence analysis of this 150-bp enhancer and identification of its multiple binding factors, including AP1, MEF2, NFAT, Runx1 and TBX5. Using this enhancer as bait, we performed yeast one-hybrid assay and identified multiple candidate Col10a1-interacting genes, including cyclooxygenase 1 (Cox-1) and Cox-2. We have also performed mass spectrometry analysis and detected EF1-alpha, Fus, GdF7 and Runx3 as components of the specific complex formed by the cis-enhancer and nuclear extracts from hypertrophic MCT (mouse chondrocytes immortalized with large T antigen) cells that express Col10a1 abundantly. Notably, some of the candidate genes are differentially expressed in hypertrophic MCT cells and have been associated with chondrocyte hypertrophy and Runx2, an indispensible Col10a1 regulator. Intriguingly, we detected high-level Cox-2 expression in

  6. Scoliosis in osteogenesis imperfecta caused by COL1A1/COL1A2 mutations - genotype-phenotype correlations and effect of bisphosphonate treatment.

    PubMed

    Sato, Atsuko; Ouellet, Jean; Muneta, Takeshi; Glorieux, Francis H; Rauch, Frank

    2016-05-01

    Bisphosphonates are widely used to treat children with osteogenesis imperfecta (OI), a bone fragility disorder that is most often caused by mutations in COL1A1 or COL1A2. However, it is unclear whether this treatment decreases the risk of developing scoliosis. We retrospectively evaluated spine radiographs and charts of 437 patients (227 female) with OI caused by mutations in COL1A1 or COL1A2 and compared the relationship between scoliosis, genotype and bisphosphonate treatment history. At the last follow-up (mean age 11.9 [SD: 5.9] years), 242 (55%) patients had scoliosis. The prevalence of scoliosis was highest in OI type III (89%), followed by OI type IV (61%) and OI type I (36%). Moderate to severe scoliosis (Cobb angle ≥25°) was rare in individuals with COL1A1 haploinsufficiency mutations but was present in about two fifth of patients with triple helical glycine substitutions or C-propeptide mutations. During the first 2 to 4years of bisphosphonate therapy, patients with OI type III had lower Cobb angle progression rates than before bisphosphonate treatment, whereas in OI types I and IV bisphosphonate treatment was not associated with a change in Cobb angle progression rates. At skeletal maturity, the prevalence of scoliosis (Cobb angle >10°) was similar in patients who had started bisphosphonate treatment early in life (before 5.0years of age) and in patients who had started therapy later (after the age of 10.0years) or had never received bisphosphonate therapy. Bisphosphonate treatment decreased progression rate of scoliosis in OI type III but there was no evidence of a positive effect on scoliosis in OI types I and IV. The prevalence of scoliosis at maturity was not influenced by the bisphosphonate treatment history in any OI type. PMID:26927310

  7. The role of mutations in COL6A3 in isolated dystonia.

    PubMed

    Lohmann, Katja; Schlicht, Felix; Svetel, Marina; Hinrichs, Frauke; Zittel, Simone; Graf, Julia; Lohnau, Thora; Schmidt, Alexander; Mir, Pablo; Krause, Patricia; Lang, Antony E; Jabusch, Hans-Christian; Wolters, Alexander; Kamm, Christoph; Zeuner, Kirsten E; Altenmüller, Eckart; Naz, Sadaf; Chung, Sun Ju; Kostic, Vladimir S; Münchau, Alexander; Kühn, Andrea A; Brüggemann, Norbert; Klein, Christine

    2016-04-01

    Specific mutations in COL6A3 have recently been reported as the cause of isolated recessive dystonia, which is a rare movement disorder. In all patients, at least one mutation was located in Exons 41 and 42. In an attempt to replicate these findings, we assessed by direct sequencing the frequency of rare variants in Exons 41 and 42 of COL6A3 in 955 patients with isolated or combined dystonia or with another movement disorder with dystonic features. We identified nine heterozygous carriers of rare variants including five different missense mutations and an extremely rare synonymous variant. In these nine patients, we sequenced the remaining 41 coding exons of COL6A3 to test for a second mutation in the compound heterozygous state. In only one of them, a second rare variant was identified (Thr732Met + Pro3082Arg). Of note, this patient had been diagnosed with Parkinson´s disease (with dystonic posturing) due to homozygous PINK1 mutations. The COL6A3 mutations clearly did not segregate with the disease in the four affected siblings of this family. Further, there was no indication for a disease-modifying effect of the COL6A3 mutations since disease severity or age at onset did not correlate with the number of COL6A3 mutated alleles in this family. In conjunction with the relatively high frequency of homozygous carriers of reported mutations in publically available databases, our data call a causal role for variants in COL6A3 in isolated dystonia into question. PMID:26872670

  8. Two different forms of lethal chondrodysplasias caused by COL2A1 gene mutations

    SciTech Connect

    Winterpacht, A.; Hilbert, K.; Schwarze, U.

    1994-09-01

    Two bone dysplasia families seem to be due to mutations in the type II procollagen gene (COL2A1): the so-called spondyloepiphyseal dysplasia congenita (SEDC) group with achondrogenesis II, hypochondrogenesis, SEDC, osteoarthrosis and the Stickler-Kniest pattern that include different forms of Kniest and Stickler dysplasia. Both groups comprise a clinical spectrum ranging from lethal to mild. COL2A1-mutations have been identified in lethal forms of the SEDC family but not in lethal forms of the Stickler/Kniest group. We now report a COL2A-1 mutation in an additional case of hypochondrogenesis (patient S) and in a lethal case of Kniest dysplasia (patient B). We amplified all 54 exons of the COL2A1 gene in both patients and screened the PCR products for mutations by SSCP analysis and sequencing. In patient B, we identified an 18 bp deletion in exon 34 which removes 6 amino acids from the mature protein. In patient S, we were able to identify a two base pair exchange (GG to AT) in exon 31, which leads to the very unusual conversion of Gly to Ile. To our knowledge, this is the first report of a Gly to Ile conversion in the COL2A1 gene, and the first report of a COL2A1 gene mutation in a lethal form of Kniest dysplasia. On the basis of the known COL2A1 gene mutations and the genotype-phenotype correlations established so far, we provide molecular data (an in frame deletion in patient B and a Gly conversion in patient S) that support their clinical classification as Kniest dysplasia and hypochondrogenesis, respectively.

  9. Epigenetic regulation of COL15A1 in smooth muscle cell replicative aging and atherosclerosis

    PubMed Central

    Connelly, Jessica J.; Cherepanova, Olga A.; Doss, Jennifer F.; Karaoli, Themistoclis; Lillard, Travis S.; Markunas, Christina A.; Nelson, Sarah; Wang, Tianyuan; Ellis, Peter D.; Langford, Cordelia F.; Haynes, Carol; Seo, David M.; Goldschmidt-Clermont, Pascal J.; Shah, Svati H.; Kraus, William E.; Hauser, Elizabeth R.; Gregory, Simon G.

    2013-01-01

    Smooth muscle cell (SMC) proliferation is a hallmark of vascular injury and disease. Global hypomethylation occurs during SMC proliferation in culture and in vivo during neointimal formation. Regardless of the programmed or stochastic nature of hypomethylation, identifying these changes is important in understanding vascular disease, as maintenance of a cells' epigenetic profile is essential for maintaining cellular phenotype. Global hypomethylation of proliferating aortic SMCs and concomitant decrease of DNMT1 expression were identified in culture during passage. An epigenome screen identified regions of the genome that were hypomethylated during proliferation and a region containing Collagen, type XV, alpha 1 (COL15A1) was selected by ‘genomic convergence’ for characterization. COL15A1 transcript and protein levels increased with passage-dependent decreases in DNA methylation and the transcript was sensitive to treatment with 5-Aza-2′-deoxycytidine, suggesting DNA methylation-mediated gene expression. Phenotypically, knockdown of COL15A1 increased SMC migration and decreased proliferation and Col15a1 expression was induced in an atherosclerotic lesion and localized to the atherosclerotic cap. A sequence variant in COL15A1 that is significantly associated with atherosclerosis (rs4142986, P = 0.017, OR = 1.434) was methylated and methylation of the risk allele correlated with decreased gene expression and increased atherosclerosis in human aorta. In summary, hypomethylation of COL15A1 occurs during SMC proliferation and the consequent increased gene expression may impact SMC phenotype and atherosclerosis formation. Hypomethylated genes, such as COL15A1, provide evidence for concomitant epigenetic regulation and genetic susceptibility, and define a class of causal targets that sit at the intersection of genetic and epigenetic predisposition in the etiology of complex disease. PMID:23912340

  10. Position of glycine substitutions in the triple helix of COL6A1, COL6A2, and COL6A3 is correlated with severity and mode of inheritance in collagen VI myopathies.

    PubMed

    Butterfield, Russell J; Foley, A Reghan; Dastgir, Jahannaz; Asman, Stephanie; Dunn, Diane M; Zou, Yaqun; Hu, Ying; Donkervoort, Sandra; Flanigan, Kevin M; Swoboda, Kathryn J; Winder, Thomas L; Weiss, Robert B; Bönnemann, Carsten G

    2013-11-01

    Glycine substitutions in the conserved Gly-X-Y motif in the triple helical (TH) domain of collagen VI are the most commonly identified mutations in the collagen VI myopathies including Ullrich congenital muscular dystrophy, Bethlem myopathy, and intermediate (INT) phenotypes. We describe clinical and genetic characteristics of 97 individuals with glycine substitutions in the TH domain of COL6A1, COL6A2, or COL6A3 and add a review of 97 published cases, for a total of 194 cases. Clinical findings include severe, INT, and mild phenotypes even from patients with identical mutations. INT phenotypes were most common, accounting for almost half of patients, emphasizing the importance of INT phenotypes to the overall phenotypic spectrum. Glycine substitutions in the TH domain are heavily clustered in a short segment N-terminal to the 17th Gly-X-Y triplet, where they are acting as dominants. The most severe cases are clustered in an even smaller region including Gly-X-Y triplets 10-15, accounting for only 5% of the TH domain. Our findings suggest that clustering of glycine substitutions in the N-terminal region of collagen VI is not based on features of the primary sequence. We hypothesize that this region may represent a functional domain within the triple helix. PMID:24038877

  11. A COL11A2 Mutation in Labrador Retrievers with Mild Disproportionate Dwarfism

    PubMed Central

    Frischknecht, Mirjam; Niehof-Oellers, Helena; Jagannathan, Vidhya; Owczarek-Lipska, Marta; Drögemüller, Cord; Dietschi, Elisabeth; Dolf, Gaudenz; Tellhelm, Bernd; Lang, Johann; Tiira, Katriina; Lohi, Hannes; Leeb, Tosso

    2013-01-01

    We describe a mild form of disproportionate dwarfism in Labrador Retrievers, which is not associated with any obvious health problems such as secondary arthrosis. We designate this phenotype as skeletal dysplasia 2 (SD2). It is inherited as a monogenic autosomal recessive trait with incomplete penetrance primarily in working lines of the Labrador Retriever breed. Using 23 cases and 37 controls we mapped the causative mutation by genome-wide association and homozygosity mapping to a 4.44 Mb interval on chromosome 12. We re-sequenced the genome of one affected dog at 30x coverage and detected 92 non-synonymous variants in the critical interval. Only two of these variants, located in the lymphotoxin A (LTA) and collagen alpha-2(XI) chain gene (COL11A2), respectively, were perfectly associated with the trait. Previously described COL11A2 variants in humans or mice lead to skeletal dysplasias and/or deafness. The dog variant associated with disproportionate dwarfism, COL11A2:c.143G>C or p.R48P, probably has only a minor effect on collagen XI function, which might explain the comparatively mild phenotype seen in our study. The identification of this candidate causative mutation thus widens the known phenotypic spectrum of COL11A2 mutations. We speculate that non-pathogenic COL11A2 variants might even contribute to the heritable variation in height. PMID:23527306

  12. Novel action of FOXL2 as mediator of Col1a2 gene autoregulation.

    PubMed

    Marongiu, Mara; Deiana, Manila; Marcia, Loredana; Sbardellati, Andrea; Asunis, Isadora; Meloni, Alessandra; Angius, Andrea; Cusano, Roberto; Loi, Angela; Crobu, Francesca; Fotia, Giorgio; Cucca, Francesco; Schlessinger, David; Crisponi, Laura

    2016-08-01

    FOXL2 belongs to the evolutionarily conserved forkhead box (FOX) superfamily and is a master transcription factor in a spectrum of developmental pathways, including ovarian and eyelid development and bone, cartilage and uterine maturation. To analyse its action, we searched for proteins that interact with FOXL2. We found that FOXL2 interacts with specific C-terminal propeptides of several fibrillary collagens. Because these propeptides can participate in feedback regulation of collagen biosynthesis, we inferred that FOXL2 could thereby affect the transcription of the cognate collagen genes. Focusing on COL1A2, we found that FOXL2 indeed affects collagen synthesis, by binding to a DNA response element located about 65Kb upstream of this gene. According to our hypothesis we found that in Foxl2(-/-) mouse ovaries, Col1a2 was elevated from birth to adulthood. The extracellular matrix (ECM) compartmentalizes the ovary during folliculogenesis, (with type I, type III and type IV collagens as primary components), and ECM composition changes during the reproductive lifespan. In Foxl2(-/-) mouse ovaries, in addition to up-regulation of Col1a2, Col3a1, Col4a1 and fibronectin were also upregulated, while laminin expression was reduced. Thus, by regulating levels of extracellular matrix components, FOXL2 may contribute to both ovarian histogenesis and the fibrosis attendant on depletion of the follicle reserve during reproductive aging and menopause. PMID:27212026

  13. COL Application Content Guide for HTGRs: Revision to RG 1.206, Part 1 - Status Report

    SciTech Connect

    Wayne Moe

    2012-08-01

    A combined license (COL) application is required by the Nuclear Regulatory Commission (NRC) for all proposed nuclear plants. The information requirements for a COL application are set forth in 10 CFR 52.79, “Contents of Applications; Technical Information in Final Safety Analysis Report.” An applicant for a modular high temperature gas-cooled reactor (HTGR) must develop and submit for NRC review and approval a COL application which conforms to these requirements. The technical information necessary to allow NRC staff to evaluate a COL application and resolve all safety issues related to a proposed nuclear plant is detailed and comprehensive. To this, Regulatory Guide (RG) 1.206, “Combined License Applications for Nuclear Power Plants” (LWR Edition), was developed to assist light water reactor (LWR) applicants in incorporating and effectively formatting required information for COL application review (Ref. 1). However, the guidance prescribed in RG 1.206 presumes a LWR design proposal consistent with the systems and functions associated with large LWR power plants currently operating under NRC license.

  14. Differential alleleic expression of the type II collagen gene (COL2A2) in osteoarthritic cartilage

    SciTech Connect

    Loughlin, J.; Irven, C.; Sykes, B.; Athanasou, N.; Carr, A.

    1995-05-01

    Osteoarthritis (OA) is a common debilitating disease resulting from the degeneration of articular cartilage. The major protein of cartilage is type II collagen, which is encoded by the COL2A1 gene. Mutations at this locus have been discovered in several individuals with inherited disorders of cartilage. We have identified 27 primary OA patients who are heterozygous for sequence dimorphisms located in the coding region of COL2A1. These dimorphisms were used to distinguish the mRNA output from each of the two COL2A1 alleles in articular cartilage obtained from each patient. Three patients demonstrated differential allelic expression and produced <12% of the normal level of mRNA from one of their COL2A1 alleles. The same allele shows reduced expression in a well-defined OA population than in a control group, suggesting the possible existence of a rare COL2A1 allele that predisposes to OA. 31 refs., 4 figs., 3 tabs.

  15. Functionally conserved cis-regulatory elements of COL18A1 identified through zebrafish transgenesis.

    PubMed

    Kague, Erika; Bessling, Seneca L; Lee, Josephine; Hu, Gui; Passos-Bueno, Maria Rita; Fisher, Shannon

    2010-01-15

    Type XVIII collagen is a component of basement membranes, and expressed prominently in the eye, blood vessels, liver, and the central nervous system. Homozygous mutations in COL18A1 lead to Knobloch Syndrome, characterized by ocular defects and occipital encephalocele. However, relatively little has been described on the role of type XVIII collagen in development, and nothing is known about the regulation of its tissue-specific expression pattern. We have used zebrafish transgenesis to identify and characterize cis-regulatory sequences controlling expression of the human gene. Candidate enhancers were selected from non-coding sequence associated with COL18A1 based on sequence conservation among mammals. Although these displayed no overt conservation with orthologous zebrafish sequences, four regions nonetheless acted as tissue-specific transcriptional enhancers in the zebrafish embryo, and together recapitulated the major aspects of col18a1 expression. Additional post-hoc computational analysis on positive enhancer sequences revealed alignments between mammalian and teleost sequences, which we hypothesize predict the corresponding zebrafish enhancers; for one of these, we demonstrate functional overlap with the orthologous human enhancer sequence. Our results provide important insight into the biological function and regulation of COL18A1, and point to additional sequences that may contribute to complex diseases involving COL18A1. More generally, we show that combining functional data with targeted analyses for phylogenetic conservation can reveal conserved cis-regulatory elements in the large number of cases where computational alignment alone falls short. PMID:19895802

  16. Association of COL1A1 polymorphism with high myopia: a Meta-analysis

    PubMed Central

    Jin, Guang-Ming; Zhao, Xiao-Jing; Chen, Ai-Ming; Chen, Yong-Xing; Li, Qin

    2016-01-01

    AIM To investigate the association between collagen type I alpha 1 (COL1A1) gene and high myopia. METHODS In this Meta-analysis, we examined 5 published case-control studies that involved 1942 high myopia cases and 2929 healthy controls to assess the association between the COL1A1 rs2075555 polymorphism and high myopia risk. We calculated the pooled odds ratios (ORs) of COL1A1 rs2075555 polymorphism in high myopia cases vs healthy controls to evaluate the strength of the association. RESULTS Overall, there was no significant difference both in the genotype and allele distributions of COL1A1 rs2075555 polymorphism between high myopia cases and healthy controls: CC vs AA OR=1.10, 95% confidence interval (CI)=0.76-1.58; AC vs AA OR=0.98, 95%CI 0.80-1.20; CC/AC vs AA/OR=1.01, 95%CI 0.84-1.22; CC vs AC/AA OR=1.06, 95%CI=0.93-1.20; C vs A OR=1.06, 95%CI 0.91-1.23). In addition, in the stratified analyses by ethnicity, no significant associations were found in any genetic model both in European and Asia cohorts. CONCLUSION Our results indicate that the COL1A1 rs2075555 polymorphism may not affect susceptibility to high myopia. PMID:27162737

  17. Detection of induced synthesis of colicin E9 using ColE9p::gfpmut2 based reporter system.

    PubMed

    Bano, Shaista; Vankemmelbeke, Mireille; Penfold, Christopher N; James, Richard

    2014-07-01

    The majority of colicin operons are regulated by an SOS response inducible promoter (SOS promoter), located at upstream of the colicin operons. Therefore, colicin synthesis is induced by DNA damaging agents like mitomycin C (MMC) because the resulting DNA damage switches on the SOS response in bacteria. In this study, we have described the strategy for fusion of the SOS promoter of the colicin E9 operon (ColE9p) with a promoterless green fluorescent reporter gene (gfpmut2). We observed that the ColE9p-gfpmut2 is inducible by MMC which confirmed that the ColE9p-gfpmut2 is sensitive to SOS response inducing agents. The data implies that the ColE9p-gfpmut2 based reporter system is suitable for monitoring the ColE9 synthesis induced by SOS response inducing agents including antibiotics. Using green fluorescent protein expression from the ColE9p-gfpmut2 as an indicator of ColE9 synthesis; we have investigated, first time, the inducing effects of cephalexin antibiotic on ColE9 synthesis. Our data demonstrated that the cephalexin has potential to induce ColE9 synthesis from E. coli JM83 host cells albeit the level of this induction is very low hence its detection required a highly sensitive method. PMID:24652519

  18. A novel COL11A1 missense mutation in siblings with non-ocular Stickler syndrome

    PubMed Central

    Kohmoto, Tomohiro; Tsuji, Atsumi; Morita, Kei-ichi; Naruto, Takuya; Masuda, Kiyoshi; Kashimada, Kenichi; Enomoto, Keisuke; Morio, Tomohiro; Harada, Hiroyuki; Imoto, Issei

    2016-01-01

    Stickler syndrome (STL) is an autosomal, dominantly inherited, clinically variable and genetically heterogeneous connective tissue disorder characterized by ocular, auditory, orofacial and skeletal abnormalities. We conducted targeted resequencing using a next-generation sequencer for molecular diagnosis of a 2-year-old girl who was clinically suspected of having STL with Pierre Robin sequence. We detected a novel heterozygous missense mutation, NM_001854.3:n.4838G>A [NM_001854.3 (COL11A1_v001):c.4520G>A], in COL11A1, resulting in a Gly to Asp substitution at position 1507 [NM_001854.3(COL11A1_i001)] within one of the collagen-like domains of the triple helical region. The same mutation was detected in her 4-year-old brother with cleft palate and high-frequency sensorineural hearing loss. PMID:27081569

  19. COL5A1 haploinsufficiency is a common molecular mechanism underlying the classical form of EDS.

    PubMed Central

    Wenstrup, R J; Florer, J B; Willing, M C; Giunta, C; Steinmann, B; Young, F; Susic, M; Cole, W G

    2000-01-01

    We have identified haploinsufficiency of the COL5A1 gene that encodes the proalpha1(V) chain of type V collagen in the classical form of the Ehlers-Danlos syndrome (EDS), a heritable connective-tissue disorder that severely alters the collagen-fibrillar structure of the dermis, joints, eyes, and blood vessels. Eight of 28 probands with classical EDS who were heterozygous for expressed polymorphisms in COL5A1 showed complete or nearly complete loss of expression of one COL5A1 allele. Reduced levels of proalpha1(V) mRNA relative to the levels of another type V collagen mRNA, proalpha2(V), were also observed in the cultured fibroblasts from EDS probands. Products of the two COL5A1 alleles were approximately equal after the addition of cycloheximide to the fibroblast cultures. After harvesting of mRNAs from cycloheximide-treated cultured fibroblasts, heteroduplex analysis of overlapping reverse transcriptase-PCR segments spanning the complete proalpha1(V) cDNA showed anomalies in four of the eight probands that led to identification of causative mutations, and, in the remaining four probands, targeting of CGA-->TGA mutations in genomic DNA revealed a premature stop at codon in one of them. We estimate that approximately one-third of individuals with classical EDS have mutations of COL5A1 that result in haploinsufficiency. These findings indicate that the normal formation of the heterotypic collagen fibrils that contain types I, III, and V collagen requires the expression of both COL5A1 alleles. PMID:10777716

  20. miRNA-29a targets COL3A1 to regulate the level of type III collagen in pig.

    PubMed

    Chuan-Hao, Li; Wei, Chen; Jia-Qing, Hu; Yan-Dong, Wang; Shou-Dong, Wang; Yong-Qing, Zeng; Hui, Wang

    2016-10-30

    COL3A1 encodes the protein, collagen type III alpha 1, which is an important component of collagen. Collagen can have a considerable effect on the processing quality of meat, and is nutritious. Bioinformatic analysis using Targetscan showed that COL3A1 could be a target gene of miRNA-29a. Moreover, we found that Laiwu pigs have higher levels of type III collagen and lower levels of miRNA-29a than Landrace pigs. Therefore, we hypothesized that miRNA-29a suppresses the expression of COL3A1 by targeting its 3'-UTR. miRNA-29a appears to play an inhibitory role in the regulation of COL3A1 in PK15 cells because of the following: (1) overexpression of miRNA-29a resulted in a significant down-regulation of COL3A1 protein levels (2) overexpression of miRNA-29a significantly decreased the level of COL3A1 mRNA. (3) The activity of a COL3A1 luciferase reporter was significant reduced by miRNA-29a. Furthermore, the levels of miRNA-29a and collagen type III in four tissues in Laiwu and Landrace pigs were consistent with the above observations. In this study, we identified COL3A1 as a direct target for miRNA-29a, which will inform further studies of meat quality. PMID:27476968

  1. A mouse model for dominant collagen VI disorders: heterozygous deletion of Col6a3 Exon 16.

    PubMed

    Pan, Te-Cheng; Zhang, Rui-Zhu; Arita, Machiko; Bogdanovich, Sasha; Adams, Sheila M; Gara, Sudheer Kumar; Wagener, Raimund; Khurana, Tejvior S; Birk, David E; Chu, Mon-Li

    2014-04-11

    Dominant and recessive mutations in collagen VI genes, COL6A1, COL6A2, and COL6A3, cause a continuous spectrum of disorders characterized by muscle weakness and connective tissue abnormalities ranging from the severe Ullrich congenital muscular dystrophy to the mild Bethlem myopathy. Herein, we report the development of a mouse model for dominant collagen VI disorders by deleting exon 16 in the Col6a3 gene. The resulting heterozygous mouse, Col6a3(+/d16), produced comparable amounts of normal Col6a3 mRNA and a mutant transcript with an in-frame deletion of 54 bp of triple-helical coding sequences, thus mimicking the most common molecular defect found in dominant Ullrich congenital muscular dystrophy patients. Biosynthetic studies of mutant fibroblasts indicated that the mutant α3(VI) collagen protein was produced and exerted a dominant-negative effect on collagen VI microfibrillar assembly. The distribution of the α3(VI)-like chains of collagen VI was not altered in mutant mice during development. The Col6a3(+/d16) mice developed histopathologic signs of myopathy and showed ultrastructural alterations of mitochondria and sarcoplasmic reticulum in muscle and abnormal collagen fibrils in tendons. The Col6a3(+/d16) mice displayed compromised muscle contractile functions and thereby provide an essential preclinical platform for developing treatment strategies for dominant collagen VI disorders. PMID:24563484

  2. A single base mutation in COL5A2 causes Ehlers-Danlos syndrome type II.

    PubMed

    Richards, A J; Martin, S; Nicholls, A C; Harrison, J B; Pope, F M; Burrows, N P

    1998-10-01

    Ehlers-Danlos syndrome (EDS) is a heterogeneous group of connective tissue disorders. Recently mutations have been found in the genes for type V collagen in a small number of people with the most common forms of EDS, types I and II. Here we characterise a COL5A2 mutation in an EDS II family. Cultured dermal fibroblasts obtained from an affected subject synthesised abnormal type V collagen. Haplotype analysis excluded COL5A1 but was concordant with COL5A2 as the disease locus. The entire open reading frame of the COL5A2 cDNA was directly sequenced and a single base mutation detected. It substituted a glycine residue within the triple helical domain (G934R) of alpha2(V) collagen, typical of the dominant negative changes in other collagens, which cause various other inherited connective tissue disorders. All three affected family members possessed the single base change, which was absent in 50 normal chromosomes. PMID:9783710

  3. Mutation Update for COL2A1 Gene Variants Associated with Type II Collagenopathies.

    PubMed

    Barat-Houari, Mouna; Sarrabay, Guillaume; Gatinois, Vincent; Fabre, Aurélie; Dumont, Bruno; Genevieve, David; Touitou, Isabelle

    2016-01-01

    Mutations in the COL2A1 gene cause a spectrum of rare autosomal-dominant conditions characterized by skeletal dysplasia, short stature, and sensorial defects. An early diagnosis is critical to providing relevant patient care and follow-up, and genetic counseling to affected families. There are no recent exhaustive descriptions of the causal mutations in the literature. Here, we provide a review of COL2A1 mutations extracted from the Leiden Open Variation Database (LOVD) that we updated with data from PubMed and our own patients. Over 700 patients were recorded, harboring 415 different mutations. One-third of the mutations are dominant-negative mutations that affect the glycine residue in the G-X-Y repeats of the alpha 1 chain. These mutations disrupt the collagen triple helix and are common in achondrogenesis type II and hypochondrogenesis. The mutations resulting in a premature stop codon are found in less severe phenotypes such as Stickler syndrome. The p.(Arg275Cys) substitution is found in all patients with COL2A1-associated Czech dysplasia. LOVD-COL2A1 provides support and potential collaborative material for scientific and clinical projects aimed at elucidating phenotype-genotype correlation and differential diagnosis in patients with type II collagenopathies. PMID:26443184

  4. Widely distributed mutations in the COL2A1 gene produce achondrogenesis type II/hypochondrogenesis.

    PubMed

    Körkkö, J; Cohn, D H; Ala-Kokko, L; Krakow, D; Prockop, D J

    2000-05-15

    The COL2A1 gene was assayed for mutations in genomic DNA from 12 patients with achondrogenesis type II/hypochondrogenesis. The exons and flanking sequences of the 54 exons in the COL2A1 gene were amplified by a series of specific primers using PCR. The PCR products were scanned for mutations by conformation sensitive gel electrophoresis, and PCR products that generated heteroduplex bands were then sequenced. Mutations in the COL2A1 gene were found in all 12 patients. Ten of the mutations were single base substitutions that converted a codon for an obligate glycine to a codon for an amino acid with a bulkier side chain. One of the mutations was a change in a consensus RNA splice site. Another was an 18-base pair deletion of coding sequences. The results confirmed previous indications that conformation sensitive gel electrophoresis is highly sensitive for detection of mutations in large and complex genes. They also demonstrate that most, if not all, patients with achondrogenesis type II/hypochondrogenesis have mutations in the COL2A1 gene. PMID:10797431

  5. Efficient detection of Alport syndrome COL4A5 mutations with multiplex genomic PCR-SSCP.

    PubMed

    Barker, D F; Denison, J C; Atkin, C L; Gregory, M C

    2001-01-15

    We have performed effective mutation screening of COL4A5 with a new method of direct, multiplex genomic amplification that employs a single buffer condition and PCR profile. Application of the method to a consecutive series of 46 United States patients with diverse indications of Alport syndrome resulted in detection of mutations in 31 cases and of five previously unreported polymorphisms. With a correction for the presence of cases that are not likely to be due to changes at the COL4A5 locus, the mutation detection sensitivity is greater than 79%. The test examines 52 segments, including the COL4A6/COL4A5 intergenic promoter region, all 51 of the previously recognized exons and two newly detected exons between exons 41 and 42 that encode an alternatively spliced mRNA segment. New genomic sequence information was generated and used to design primer pairs that span substantial intron sequences on each side of all 53 exons. For SSCP screening, 16 multiplex PCR combinations (15 4-plex and 1 3-plex) were used to provide complete, partially redundant coverage of the gene. The selected combinations allow clear resolution of products from each segment using various SSCP gel formulations. One of the 29 different mutations detected initially seemed to be a missense change in exon 32 but was found to cause exon skipping. Another missense variant may mark a novel functional site located in the collagenous domain. PMID:11223851

  6. Fibrochondrogenesis Results from Mutations in the COL11A1 Type XI Collagen Gene

    PubMed Central

    Tompson, Stuart W.; Bacino, Carlos A.; Safina, Nicole P.; Bober, Michael B.; Proud, Virginia K.; Funari, Tara; Wangler, Michael F.; Nevarez, Lisette; Ala-Kokko, Leena; Wilcox, William R.; Eyre, David R.; Krakow, Deborah; Cohn, Daniel H.

    2010-01-01

    Fibrochondrogenesis is a severe, autosomal-recessive, short-limbed skeletal dysplasia. In a single case of fibrochondrogenesis, whole-genome SNP genotyping identified unknown ancestral consanguinity by detecting three autozygous regions. Because of the predominantly skeletal nature of the phenotype, the 389 genes localized to the autozygous intervals were prioritized for mutation analysis by correlation of their expression with known cartilage-selective genes via the UCLA Gene Expression Tool, UGET. The gene encoding the α1 chain of type XI collagen (COL11A1) was the only cartilage-selective gene among the three candidate intervals. Sequence analysis of COL11A1 in two genetically independent fibrochondrogenesis cases demonstrated that each was a compound heterozygote for a loss-of-function mutation on one allele and a mutation predicting substitution for a conserved triple-helical glycine residue on the other. The parents who were carriers of missense mutations had myopia. Early-onset hearing loss was noted in both parents who carried a loss-of-function allele, suggesting COL11A1 as a locus for mild, dominantly inherited hearing loss. These findings identify COL11A1 as a locus for fibrochondrogenesis and indicate that there might be phenotypic manifestations among carriers. PMID:21035103

  7. Fibrochondrogenesis results from mutations in the COL11A1 type XI collagen gene.

    PubMed

    Tompson, Stuart W; Bacino, Carlos A; Safina, Nicole P; Bober, Michael B; Proud, Virginia K; Funari, Tara; Wangler, Michael F; Nevarez, Lisette; Ala-Kokko, Leena; Wilcox, William R; Eyre, David R; Krakow, Deborah; Cohn, Daniel H

    2010-11-12

    Fibrochondrogenesis is a severe, autosomal-recessive, short-limbed skeletal dysplasia. In a single case of fibrochondrogenesis, whole-genome SNP genotyping identified unknown ancestral consanguinity by detecting three autozygous regions. Because of the predominantly skeletal nature of the phenotype, the 389 genes localized to the autozygous intervals were prioritized for mutation analysis by correlation of their expression with known cartilage-selective genes via the UCLA Gene Expression Tool, UGET. The gene encoding the α1 chain of type XI collagen (COL11A1) was the only cartilage-selective gene among the three candidate intervals. Sequence analysis of COL11A1 in two genetically independent fibrochondrogenesis cases demonstrated that each was a compound heterozygote for a loss-of-function mutation on one allele and a mutation predicting substitution for a conserved triple-helical glycine residue on the other. The parents who were carriers of missense mutations had myopia. Early-onset hearing loss was noted in both parents who carried a loss-of-function allele, suggesting COL11A1 as a locus for mild, dominantly inherited hearing loss. These findings identify COL11A1 as a locus for fibrochondrogenesis and indicate that there might be phenotypic manifestations among carriers. PMID:21035103

  8. Why deep drilling in the Colônia Basin (Brazil)?

    NASA Astrophysics Data System (ADS)

    Ledru, M.-P.; Reimold, W. U.; Ariztegui, D.; Bard, E.; Crósta, A. P.; Riccomini, C.; Sawakuchi, A. O.

    2015-12-01

    The Colônia Deep Drilling Project held its first International Continental Scientific Drilling Program (ICDP) workshop in September 2014 at the University of São Paulo (Brazil). Twenty-seven experts from six countries discussed the feasibility and the expectations of a deep drilling in the structure of Colônia located at the southwestern margin of the city of São Paulo. After presenting the studies performed at the site during the last decades, participants focused on the objectives, priorities and detailed planning for a full deep-drilling proposal. An excursion to the site and new auger coring showed the importance of the Colônia site for studying the evolution of a tropical rainforest and to evaluate the interplay between the South American summer monsoon, the Intertropical Convergence Zone (ITCZ) and the southern Westerlies belt during the last 5 million years. In addition, deep drilling will eventually solve the still unresolved issue of the origin of the structure of Colônia as a result of meteorite impact or endogenous processes.

  9. A rapid and efficient newly established method to detect COL1A1-PDGFB gene fusion in dermatofibrosarcoma protuberans

    SciTech Connect

    Yokoyama, Yoko; Shimizu, Akira; Okada, Etsuko; Ishikawa, Osamu; Motegi, Sei-ichiro

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer We developed new method to rapidly identify COL1A1-PDGFB fusion in DFSP. Black-Right-Pointing-Pointer New PCR method using a single primer pair detected COL1A1-PDGFB fusion in DFSP. Black-Right-Pointing-Pointer This is the first report of DFSP with a novel COL1A1 breakpoint in exon 5. -- Abstract: The detection of fusion transcripts of the collagen type 1{alpha}1 (COL1A1) and platelet-derived growth factor-BB (PDGFB) genes by genetic analysis has recognized as a reliable and valuable molecular tool for the diagnosis of dermatofibrosarcoma protuberans (DFSP). To detect the COL1A1-PDGFB fusion, almost previous reports performed reverse transcription polymerase chain reaction (RT-PCR) using multiplex forward primers from COL1A1. However, it has possible technical difficulties with respect to the handling of multiple primers and reagents in the procedure. The objective of this study is to establish a rapid, easy, and efficient one-step method of PCR using only a single primer pair to detect the fusion transcripts of the COL1A1 and PDGFB in DFSP. To validate new method, we compared the results of RT-PCR in five patients of DFSP between the previous method using multiplex primers and our established one-step RT-PCR using a single primer pair. In all cases of DFSP, the COL1A1-PDGFB fusion was detected by both previous method and newly established one-step PCR. Importantly, we detected a novel COL1A1 breakpoint in exon 5. The newly developed method is valuable to rapidly identify COL1A1-PDGFB fusion transcripts in DFSP.

  10. Collagen XXIV (Col24α1) Promotes Osteoblastic Differentiation and Mineralization through TGF-β/Smads Signaling Pathway

    PubMed Central

    Wang, Weizhuo; Olson, Douglas; Liang, Gang; Franceschi, Renny T; Li, Chunyi; Wang, Bingyan; Wang, Shuen Shiuan; Yang, Shuying

    2012-01-01

    Collagen XXIV (Col24α1) is a recently discovered fibrillar collagen. It is known that mouse Col24α1 is predominantly expressed in the forming skeleton of the mouse embryo, as well as in the trabecular bone and periosteum of the newborn mouse. However, the role and mechanism of Col24α1 in osteoblast differentiation and mineralization remains unclear. By analyzing the expression pattern of Col24α1, we confirmed that it is primarily expressed in bone tissues, and this expression gradually increased concomitant with the progression of osteoblast differentiation. Through the use of a lentivirus vector-mediated interference system, silencing Col24α1 expression in MC3T3-E1 murine preosteoblastic cells resulted in significant inhibition of alkaline phosphatase (ALP) activity, cell mineralization, and the expression of osteoblast marker genes such as runt-related transcription factor 2 (Runx2), osteocalcin (OCN), ALP, and type I collagen (Col I). Subsequent overexpression not only rescued the deficiency in osteoblast differentiation from Col24α1 silenced cells, but also enhanced osteoblastic differentiation in control cells. We further revealed that Col24α1 interacts with integrin β3, and silencing Col24α1 up-regulated the expression of Smad7 during osteoblast differentiation while at the same time inhibiting the phosphorylation of the Smad2/3 complex. These results suggest that Col24α1 imparts some of its regulatory control on osteoblast differentiation and mineralization at least partially through interaction with integrin β3 and the transforming growth factor beta (TGF-β) /Smads signaling pathway. PMID:23139630

  11. Familial Ehlers-Danlos syndrome with lethal arterial events caused by a mutation in COL5A1.

    PubMed

    Monroe, Glen R; Harakalova, Magdalena; van der Crabben, Saskia N; Majoor-Krakauer, Danielle; Bertoli-Avella, Aida M; Moll, Frans L; Oranen, Björn I; Dooijes, Dennis; Vink, Aryan; Knoers, Nine V; Maugeri, Alessandra; Pals, Gerard; Nijman, Isaac J; van Haaften, Gijs; Baas, Annette F

    2015-06-01

    Different forms of Ehlers-Danlos syndrome (EDS) exist, with specific phenotypes and associated genes. Vascular EDS, caused by heterozygous mutations in the COL3A1 gene, is characterized by fragile vasculature with a high risk of catastrophic vascular events at a young age. Classic EDS, caused by heterozygous mutations in the COL5A1 or COL5A2 genes, is characterized by fragile, hyperextensible skin and joint laxity. To date, vessel rupture in four unrelated classic EDS patients with a confirmed COL5A1 mutation has been reported. We describe familial occurrence of a phenotype resembling vascular EDS in a mother and her two sons, who all died at an early age from arterial ruptures. Diagnostic Sanger sequencing in the proband failed to detect aberrations in COL3A1, COL1A1, COL1A2, TGFBR1, TGFBR2, SMAD3, and ACTA2. Next, the proband's DNA was analyzed using a next-generation sequencing approach targeting 554 genes linked to vascular disease (VASCULOME project). A novel heterozygous mutation in COL5A1 was detected, resulting in an essential glycine substitution at the C-terminal end of the triple helix domain (NM_000093.4:c.4610G>T; p.Gly1537Val). This mutation was also present in DNA isolated from autopsy material of the index's brother. No material was available from the mother, but the mutation was excluded in her parents, siblings and in the father of her sons, suggesting that the COL5A1 mutation occurred in the mother's genome de novo. In conclusion, we report familial occurrence of lethal arterial events caused by a COL5A1 mutation. PMID:25845371

  12. Parental allelic variation at COL6A1 and congenital heart defects in trisomy 21

    SciTech Connect

    Kessling, A.M.; Howard, C.M.; Farrer, M.J.

    1994-09-01

    Overt congenital heart defects (CHD) affect over 40% of newborns with Down syndrome. On the hypothesis that genetic variation on chromosome 21 determines this clinical variability, we studied a CHD candidate locus (COL6A1) on 21q22.3. We studied three RFLP loci in COL6A1 in 37 families of known British/Irish population of ancestral origin, and in population-matched controls. Each family had a child with trisomy 21 with or without accompanying congenital heart defect (CHD). Parental and meiotic origin of nondisjunction were determined using peri-centromeric markers. For the analysis, we considered groups of families with trisomic children with and without CHD, and subsets of nondisjoining and disjoining parents. Parental genotypes at nine control RFLP loci on chromosome 21 showed no association with CHD in the trisomic child. By contrast, parental genotypes at all three individual RFLP loci within COL6A1 showed statistically significant association with the trisomic child`s CHD status. Pairwise consideration of these loci in groups of families of trisomic children with and without CHD showed subsets of nondisjoining and disjoining parents to have different linkage disequilibrium patterns at these loci than population-matched controls. This suggests that the COL6A1 alleles of the parents are not representative of the population as a whole. Consideration of all three loci together as haplotypes supports this conclusion. Four results suggest that a functional mutation within, or in linkage disequilibrium with COL6A1 influences CHD outcome in trisomy 21.

  13. Stroma derived COL6A3 is a potential prognosis marker of colorectal carcinoma revealed by quantitative proteomics

    PubMed Central

    Chen, Sun-Xia; Wang, Xiao-Qing; Cui, Shu-Jian; Liu, Xiao-Hui; Jiang, Ying-Hua; Wang, Jie; Zhang, Yang; Yang, Peng-Yuan; Liu, Feng

    2015-01-01

    Colorectal cancer (CRC) represents the third most common cancer in males and second in females worldwide. Here, we performed a quantitative 8-plex iTRAQ proteomics analysis of the secreted proteins from five colonic fibroblast cultures and three colon cancer epithelial cell lines. We identified 1114 proteins at 0% FDR, including 587 potential secreted proteins. We further recognized 116 fibroblast-enriched proteins which were significantly associated with cell movement, angiogenesis, proliferation and wound healing, and 44 epithelial cell-enriched proteins. By interrogation of Oncomine database, we found that 20 and 8 fibroblast-enriched proteins were up- and downregulated in CRC, respectively. Western blots confirmed the fibroblast-specific secretion of filamin C, COL6A3, COL4A1 and spondin-2. Upregulated mRNA and stroma expression of COL6A3 in CRC, which were revealed by Oncomine analyses and tissue-microarray-immunohistochemistry, indicated poor prognosis. COL6A3 expression was significantly associated with Dukes stage, T stage, stage, recurrence and smoking status. Circulating plasma COL6A3 in CRC patients was upregulated significantly comparing with healthy peoples. Receiver operating characteristic curve analysis revealed that COL6A3 has better predictive performance for CRC with an area under the curve of 0.885 and the best sensitivity/specificity of 92.9%/81.3%. Thus we demonstrated that COL6A3 was a potential diagnosis and prognosis marker of CRC. PMID:26338966

  14. Stroma derived COL6A3 is a potential prognosis marker of colorectal carcinoma revealed by quantitative proteomics.

    PubMed

    Qiao, Jie; Fang, Cai-Yun; Chen, Sun-Xia; Wang, Xiao-Qing; Cui, Shu-Jian; Liu, Xiao-Hui; Jiang, Ying-Hua; Wang, Jie; Zhang, Yang; Yang, Peng-Yuan; Liu, Feng

    2015-10-01

    Colorectal cancer (CRC) represents the third most common cancer in males and second in females worldwide. Here, we performed a quantitative 8-plex iTRAQ proteomics analysis of the secreted proteins from five colonic fibroblast cultures and three colon cancer epithelial cell lines. We identified 1114 proteins at 0% FDR, including 587 potential secreted proteins. We further recognized 116 fibroblast-enriched proteins which were significantly associated with cell movement, angiogenesis, proliferation and wound healing, and 44 epithelial cell-enriched proteins. By interrogation of Oncomine database, we found that 20 and 8 fibroblast-enriched proteins were up- and downregulated in CRC, respectively. Western blots confirmed the fibroblast-specific secretion of filamin C, COL6A3, COL4A1 and spondin-2. Upregulated mRNA and stroma expression of COL6A3 in CRC, which were revealed by Oncomine analyses and tissue-microarray-immunohistochemistry, indicated poor prognosis. COL6A3 expression was significantly associated with Dukes stage, T stage, stage, recurrence and smoking status. Circulating plasma COL6A3 in CRC patients was upregulated significantly comparing with healthy peoples. Receiver operating characteristic curve analysis revealed that COL6A3 has better predictive performance for CRC with an area under the curve of 0.885 and the best sensitivity/specificity of 92.9%/81.3%. Thus we demonstrated that COL6A3 was a potential diagnosis and prognosis marker of CRC. PMID:26338966

  15. Molecular characterization and expression profiles of MaCOL1, a CONSTANS-like gene in banana fruit.

    PubMed

    Chen, Jiao; Chen, Jian-Ye; Wang, Jun-Ning; Kuang, Jian-Fei; Shan, Wei; Lu, Wang-Jin

    2012-04-01

    CONSTANS (CO) gene is a key transcription regulator that controls the long-day induction of flowering in Arabidopsis plant. However, CO gene involved in fruit ripening and stress responses is poorly understood. In the present study, a novel cDNA encoding CONSTANS-like gene, designated as MaCOL1 was isolated and characterized from banana fruit. The full length cDNA sequence was 1887bp with an open reading frame (ORF) of 1242bp, encoding 414 amino acids with a molecular weight of 46.20kDa and a theoretical isoelectric point of 5.40. Sequence alignment showed that MaCOL1 contained two B-box zinc finger motifs and a CCT domain. In addition, MaCOL1 showed transcriptional activity in yeast and was a nucleus-localized protein. Real-time PCR analysis showed that MaCOL1 was differentially expressed among various banana plant organs, with higher expression in flower. Expression of MaCOL1 in peel changed slightly, while accumulation of MaCOL1 transcripts in pulp obviously increased during natural or ethylene-induced fruit ripening, suggesting that MaCOL1 might be associated with the pulp ripening of banana fruit. Moreover, accumulation of MaCOL1 transcript was obviously enhanced by abiotic and biotic stresses, such as chilling and pathogen Colletotrichum musae infection. Taken together, our results suggest that MaCOL1 is a transcription activator and may be involved in fruit ripening and stress responses. PMID:22285923

  16. Evidence for large superhumps in TX Col and V4742 Sgr

    NASA Astrophysics Data System (ADS)

    Retter, Alon; Liu, Alexander; Bos, Marc

    Since the discovery of the largest positive superhump period in TV Col (6.4 h), we have started a program to search for superhumps in cataclysmic variables (CVs) with large orbital periods. In this work, we summarize preliminary results of our observations of TX Col and V4742 Sgr. TX Col is an intermediate polar with a 5.7-h orbital period. V4742 Sgr is a recent (2002) nova with no known periods. CCD unfiltered continuous photometry of these two objects was carried out during 56 nights (350 hours) in 2002-2003. The time series analysis reveals the presence of several periods in both power spectra. In TX Col, in addition to the orbital period of 5.7 h, we found peaks at 7.1 h and 5.0 h. These are interpreted as positive and negative superhumps correspondingly, although the effects of the quasi-periodic oscillations at ~2 h (which may cause spurious signals) were not taken into consideration. In the light curve of V4742 Sgr two long periods are detected - 6.1 and 5.4 h as well as a short-term period at 1.6 h. This result suggests that V4742 Sgr is an intermediate polar candidate and a permanent superhump system with a large orbital period (5.4 h) and a superhump period excess of 13%. If these results are confirmed, TX Col and V4742 Sgr join TV Col to form a group of intermediate polars with extremely large superhump periods. There seems to be now growing evidence that superhumps can occur in intermediate polars with long orbital periods, which is very likely inconsistent with the theoretical prediction that superhumps can only occur in systems with mass ratios below 0.33. Alternatively, if the mass ratio in these systems is nevertheless below the theoretical limit, they should harbour undermassive secondaries and very massive white dwarfs, near the Chandrasekhar limit, which would make them excellent candidates for progenitors of supernovae type Ia.

  17. Synergistic interaction of platelet derived growth factor (PDGF) with the surface of PLLA/Col/HA and PLLA/HA scaffolds produces rapid osteogenic differentiation.

    PubMed

    Raghavendran, Hanumantha Rao Balaji; Mohan, Saktiswaren; Genasan, Krishnamurithy; Murali, Malliga Raman; Naveen, Sangeetha Vasudevaraj; Talebian, Sepehr; McKean, Robert; Kamarul, Tunku

    2016-03-01

    Scaffolds with structural features similar to the extracellular matrix stimulate rapid osteogenic differentiation in favorable microenvironment and with growth factor supplementation. In this study, the osteogenic potential of electrospun poly-l-lactide/hydroxyapatite/collagen (PLLA/Col/HA, PLLA/HA and PLLA/Col) scaffolds were tested in vitro with the supplementation of platelet derived growth factor-BB (PDGF-BB). Cell attachment and topography, mineralization, extracellular matrix protein localization, and gene expression of the human mesenchymal stromal cells were compared between the fibrous scaffolds PLLA/Col/HA, PLLA/Col, and PLLA/HA. The levels of osteocalcin, calcium, and mineralization were significantly greater in the PLLA/Col/HA and PLLA/HA compared with PLLA/Col. High expression of fibronectin, intracellular adhesion molecule, cadherin, and collagen 1 (Col1) suggests that PLLA/Col/HA and PLLA/HA scaffolds had superior osteoinductivity than PLLA/Col. Additionally, osteopontin, osteocalcin, osterix, Runt-related transcription factor 2 (Runx2), and bone morphogenic protein (BMP2) expression were higher in PLLA/Col/HA and PLLA/HA compared with PLLA/Col. In comparison with PLLA/Col, the PLLA/Col/HA and PLLA/HA scaffolds presented a significant upregulation of the genes Runx2, Col 1, Integrin, osteonectin (ON), bone gamma-carboxyglutamic acid-containing protein (BGALP), osteopontin (OPN), and BMP2. The upregulation of these genes was further increased with PDGF-BB supplementation. These results show that PDGF-BB acts synergistically with PLLA/Col/HA and PLLA/HA to enhance the osteogenic differentiation potential. Therefore, this combination can be used for the rapid expansion of bone marrow stromal cells into bone-forming cells for tissue engineering. PMID:26700235

  18. Bone mineral properties in growing Col1a2(+/G610C) mice, an animal model of osteogenesis imperfecta.

    PubMed

    Masci, Marco; Wang, Min; Imbert, Laurianne; Barnes, Aileen M; Spevak, Lyudmila; Lukashova, Lyudmila; Huang, Yihe; Ma, Yan; Marini, Joan C; Jacobsen, Christina M; Warman, Matthew L; Boskey, Adele L

    2016-06-01

    The Col1a2(+/G610C) knock-in mouse, models osteogenesis imperfecta in a large old order Amish family (OOA) with type IV OI, caused by a G-to-T transversion at nucleotide 2098, which alters the gly-610 codon in the triple-helical domain of the α2(I) chain of type I collagen. Mineral and matrix properties of the long bones and vertebrae of male Col1a2(+/G610C) and their wild-type controls (Col1a2(+/+)), were characterized to gain insight into the role of α2-chain collagen mutations in mineralization. Additionally, we examined the rescuability of the composition by sclerostin inhibition initiated by crossing Col1a2(+/G610C) with an LRP(+/A214V) high bone mass allele. At age 10-days, vertebrae and tibia showed few alterations by micro-CT or Fourier transform infrared imaging (FTIRI). At 2-months-of-age, Col1a2(+/G610C) tibias had 13% fewer secondary trabeculae than Col1a2(+/+), these were thinner (11%) and more widely spaced (20%) than those of Col1a2(+/+) mice. Vertebrae of Col1a2(+/G610C) mice at 2-months also had lower bone volume fraction (38%), trabecular number (13%), thickness (13%) and connectivity density (32%) compared to Col1(a2+/+). The cortical bone of Col1a2(+/G610C) tibias at 2-months had 3% higher tissue mineral density compared to Col1a2(+/+); Col1a2(+/G610C) vertebrae had lower cortical thickness (29%), bone area (37%) and polar moment of inertia (38%) relative to Col1a2(+/+). FTIRI analysis, which provides information on bone chemical composition at ~7μm-spatial resolution, showed tibias at 10-days did not differ between genotypes. Comparing identical bone types in Col1a2(+/G610C) to Col1a2(+/+) at 2-months-of-age, tibias showed higher mineral-to-matrix ratio in trabeculae (17%) and cortices (31%). and in vertebral cortices (28%). Collagen maturity was 42% higher at 10-days-of-age in Col1a2(+/G610C) vertebral trabeculae and in 2-month tibial cortices (12%), vertebral trabeculae (42%) and vertebral cortices (12%). Higher acid-phosphate substitution

  19. COL1A1 and COL1A2 sequencing results in cohort of patients undergoing evaluation for potential child abuse.

    PubMed

    Zarate, Yuri A; Clingenpeel, Rachel; Sellars, Elizabeth A; Tang, Xinyu; Kaylor, Julie A; Bosanko, Katherine; Linam, Leann E; Byers, Peter H

    2016-07-01

    Child abuse is a major public health concern that can explain a proportion of fractures in children. Osteogenesis imperfecta (OI) is the most common inherited syndrome that predisposes to skeletal fractures. We conducted a retrospective analysis of data from clinical, laboratory, and radiographic information from children evaluated for child abuse in which molecular testing for COL1A1 and COL1A2 genes was conducted. A total of 43 patients underwent molecular testing for OI. Pathogenic variants predicted to result in a mild form of OI were found in two patients (5%), both clinically suspected to have this diagnosis. None of the cases in whom OI molecular testing was ordered when maltreatment concerns were thought to be more likely (0/35) were identified to have pathogenic variants. After reviewing each individual case, the final diagnosis was child abuse for 34 cases (77%), and additional radiographic and laboratory studies did not identify any with inherited metabolic predisposition to fracture or rickets. We conclude that routine testing for OI in the setting of child abuse when no other suggestive clinical findings are present has a low yield. A careful review of the medical history and a detailed clinical evaluation help identify those at risk for genetic alterations. © 2016 Wiley Periodicals, Inc. PMID:27090748

  20. Somatic mosaicism and the phenotypic expression of COL2A1 mutations.

    PubMed

    Nagendran, Sonali; Richards, Allan J; McNinch, Annie; Sandford, Richard N; Snead, Martin P

    2012-05-01

    Mutations in COL2A1, the gene for type II-collagen, can result in a wide variety of phenotypes depending upon the nature of the mutation. Dominant negative mutations tend to result in severe and often lethal skeletal dysplasias such as achondrogenesis type 2, Kniest dysplasia, and spondyloepiphyseal dysplasia congenita. Stickler syndrome, a condition characterized by ophthalmological and orofacial features, deafness and arthritis, usually, but not exclusively, results from haploinsufficiency. Overlapping features of all these disorders can also be seen in the same family. Rare reports have demonstrated that phenotypic variability can be explained in some families by somatic mosaicism. Here, we describe five further examples of somatic mosaicism of COL2A1 mutations illustrating the importance of detailed clinical evaluation and molecular testing even in clinically normal parents of affected individuals. PMID:22496037

  1. Identification of mutations in the COL4A5 collagen gene in Alport syndrome.

    PubMed

    Barker, D F; Hostikka, S L; Zhou, J; Chow, L T; Oliphant, A R; Gerken, S C; Gregory, M C; Skolnick, M H; Atkin, C L; Tryggvason, K

    1990-06-01

    X-linked Alport syndrome is a hereditary glomerulonephritis in which progressive loss of kidney function is often accompanied by progressive loss of hearing. Ultrastructural defects in glomerular basement membranes (GBM) of Alport syndrome patients implicate an altered structural protein as the cause of nephritis. The product of COL4A5, the alpha 5(IV) collagen chain, is a specific component of GBM within the kidney, and the gene maps to the same X chromosomal region as does Alport syndrome. Three structural aberrations were found in COL4A5, in intragenic deletion, a Pst I site variant, and an uncharacterized abnormality, which appear to cause nephritis and deafness, with allele-specific severity, in three Alport syndrome kindreds in Utah. PMID:2349482

  2. Autosomal dominant (Beukes) premature degenerative osteoarthropathy of the hip joint unlinked to COL2A1

    SciTech Connect

    Beighton, P.; Ramesar, R.; Cilliers, H.J.

    1994-12-01

    Molecular investigations have been undertaken in several separate large South African families with autosomal dominant skeletal dysplasias in which premature degenerative osteoarthropathy of the hip joint was the major manifestation. There are sometimes additional minor changes in the spine and these conditions fall into the general spondyloepiphyseal dysplasia (SED) nosological category. In some kindreds, linkage between phenotype and the type II collagen gene (COL2A1) has been established, while in others there is no linkage. We have now completed molecular linkage investigations in an Afrikaner family named Beukes, in which 47 members in 6 generations have premature osteoarthropathy of the hip joint. A LOD score of minus infinity indicates that this condition is not the result of a defect of the COL2A1 gene. 12 refs., 2 figs., 1 tab.

  3. Structural constraints on the evolution of the collagen fibril: convergence on a 1014-residue COL domain

    PubMed Central

    Slatter, David Anthony; Farndale, Richard William

    2015-01-01

    Type I collagen is the fundamental component of the extracellular matrix. Its α1 gene is the direct descendant of ancestral fibrillar collagen and contains 57 exons encoding the rod-like triple-helical COL domain. We trace the evolution of the COL domain from a primordial collagen 18 residues in length to its present 1014 residues, the limit of its possible length. In order to maintain and improve the essential structural features of collagen during evolution, exons can be added or extended only in permitted, non-random increments that preserve the position of spatially sensitive cross-linkage sites. Such sites cannot be maintained unless the twist of the triple helix is close to 30 amino acids per turn. Inspection of the gene structure of other long structural proteins, fibronectin and titin, suggests that their evolution might have been subject to similar constraints. PMID:25994354

  4. High Himalayan meteorology: Weather at the South Col of Mount Everest

    NASA Astrophysics Data System (ADS)

    Moore, G. W. K.; Semple, John L.

    2004-09-01

    Mount Everest is often referred to as the earth's `third' pole. As such it is relatively inaccessible and little is known about its meteorology. In 1998, a portable weather station was operated at the mountain's South Col, elevation 7,986 m. We believe that this represents the highest elevation at which continuous weather data has ever been collected and thus represents a valuable dataset with which to investigate the meteorology of the high Himalaya. In this paper, we compare the observations with reanalyses from the National Centers for Environmental Prediction and the European Center for Medium-Range Weather Forecasts. We find that both reanalyses capture much of the synoptic-scale variability in the temperature and pressure at the South Col site, especially in the pre-monsoon season. Furthermore, we show that an observed weather event was the result of convection associated with a jet streak and an intrusion of stratospheric air into the upper-troposphere.

  5. A Nonsense Variant in COL6A1 in Landseer Dogs with Muscular Dystrophy.

    PubMed

    Steffen, Frank; Bilzer, Thomas; Brands, Jan; Golini, Lorenzo; Jagannathan, Vidhya; Wiedmer, Michaela; Drögemüller, Michaela; Drögemüller, Cord; Leeb, Tosso

    2015-12-01

    A novel canine muscular dystrophy in Landseer dogs was observed. We had access to five affected dogs from two litters. The clinical signs started at a few weeks of age, and the severe progressive muscle weakness led to euthanasia between 5 and 15 months of age. The pedigrees of the affected dogs suggested a monogenic autosomal-recessive inheritance of the trait. Linkage and homozygosity mapping indicated two potential genome segments for the causative variant on chromosomes 10 and 31 harboring a total of 4.8 Mb of DNA or 0.2% of the canine genome. Using the Illumina sequencing technology, we obtained a whole-genome sequence from one affected Landseer. Variants were called with respect to the dog reference genome and compared with the genetic variants of 170 control dogs from other breeds. The affected Landseer dog was homozygous for a single, private nonsynonymous variant in the critical intervals, a nonsense variant in the COL6A1 gene (Chr31:39,303,964G>T; COL6A1:c.289G>T; p.E97*). Genotypes at this variant showed perfect concordance with the muscular dystrophy phenotype in all five cases and more than 1000 control dogs. Variants in the human COL6A1 gene cause Bethlem myopathy or Ullrich congenital muscular dystrophy. We therefore conclude that the identified canine COL6A1 variant is most likely causative for the observed muscular dystrophy in Landseer dogs. On the basis of the nature of the genetic variant in Landseer dogs and their severe clinical phenotype these dogs represent a model for human Ullrich congenital muscular dystrophy. PMID:26438297

  6. A Nonsense Variant in COL6A1 in Landseer Dogs with Muscular Dystrophy

    PubMed Central

    Steffen, Frank; Bilzer, Thomas; Brands, Jan; Golini, Lorenzo; Jagannathan, Vidhya; Wiedmer, Michaela; Drögemüller, Michaela; Drögemüller, Cord; Leeb, Tosso

    2015-01-01

    A novel canine muscular dystrophy in Landseer dogs was observed. We had access to five affected dogs from two litters. The clinical signs started at a few weeks of age, and the severe progressive muscle weakness led to euthanasia between 5 and 15 months of age. The pedigrees of the affected dogs suggested a monogenic autosomal-recessive inheritance of the trait. Linkage and homozygosity mapping indicated two potential genome segments for the causative variant on chromosomes 10 and 31 harboring a total of 4.8 Mb of DNA or 0.2% of the canine genome. Using the Illumina sequencing technology, we obtained a whole-genome sequence from one affected Landseer. Variants were called with respect to the dog reference genome and compared with the genetic variants of 170 control dogs from other breeds. The affected Landseer dog was homozygous for a single, private nonsynonymous variant in the critical intervals, a nonsense variant in the COL6A1 gene (Chr31:39,303,964G>T; COL6A1:c.289G>T; p.E97*). Genotypes at this variant showed perfect concordance with the muscular dystrophy phenotype in all five cases and more than 1000 control dogs. Variants in the human COL6A1 gene cause Bethlem myopathy or Ullrich congenital muscular dystrophy. We therefore conclude that the identified canine COL6A1 variant is most likely causative for the observed muscular dystrophy in Landseer dogs. On the basis of the nature of the genetic variant in Landseer dogs and their severe clinical phenotype these dogs represent a model for human Ullrich congenital muscular dystrophy. PMID:26438297

  7. A COL7A1 Mutation Causes Dystrophic Epidermolysis Bullosa in Rotes Höhenvieh Cattle

    PubMed Central

    Menoud, Annie; Welle, Monika; Tetens, Jens; Lichtner, Peter; Drögemüller, Cord

    2012-01-01

    We identified a congenital mechanobullous skin disorder in six calves on a single farm of an endangered German cattle breed in 2010. The condition presented as a large loss of skin distal to the fetlocks and at the mucosa of the muzzle. All affected calves were euthanized on humane grounds due to the severity, extent and progression of the skin and oral lesions. Examination of skin samples under light microscopy revealed detachment of the epidermis from the dermis at the level of the dermo epidermal junction, leading to the diagnosis of a subepidermal bullous dermatosis such as epidermolysis bullosa. The pedigree was consistent with monogenic autosomal recessive inheritance. We localized the causative mutation to an 18 Mb interval on chromosome 22 by homozygosity mapping. The COL7A1 gene encoding collagen type VII alpha 1 is located within this interval and COL7A1 mutations have been shown to cause inherited dystrophic epidermolysis bullosa (DEB) in humans. A SNP in the bovine COL7A1 exon 49 (c.4756C>T) was perfectly associated with the observed disease. The homozygous mutant T/T genotype was exclusively present in affected calves and their parents were heterozygous C/T confirming the assumed recessive mode of inheritance. All known cases and genotyped carriers were related to a single cow, which is supposed to be the founder animal. The mutant T allele was absent in 63 animals from 24 cattle breeds. The identified mutation causes a premature stop codon which leads to a truncated protein representing a complete loss of COL7A1 function (p.R1586*). We thus have identified a candidate causative mutation for this genetic disease using only three cases to unravel its molecular basis. Selection against this mutation can now be used to eliminate the mutant allele from the Rotes Höhenvieh breed. PMID:22715415

  8. Transcriptional promoter of the human alpha 1(V) collagen gene (COL5A1).

    PubMed Central

    Lee, S; Greenspan, D S

    1995-01-01

    We have characterized the 5' region of the human alpha 1(V) collagen gene (COL5A1). The transcriptional promoter is shown to have a number of features characteristic of the promoters of 'housekeeping' and growth-control-related genes. It lacks obvious TATA and CAAT boxes, has multiple transcription start sites, has a high GC content, lies within a well-defined CpG island and has a number of consensus sites for the potential binding of transcription factor Sp1. This type of promoter structure, while unusual for a collagen gene, is consistent with the broad distribution of expression of COL5A1 and is reminiscent of the promoter structures of the genes encoding type VI collagen, which has a similarly broad distribution of expression. Stepwise deletion of COL5A1 5' sequences, placed upstream of a heterologous reporter gene, yielded a gradual decrease in promoter activity, indicating that the COL5A1 promoter is composed of an array of cis-acting elements. A minimal promoter region contained within the 212 bp immediately upstream of the major transcription start site contained no consensus sequences for the binding of known transcription factors, but gel mobility shift assays showed this region to bind nuclear factors, including Sp1, at a number of sites. The major transcription start site is flanked by an upstream 34-bp oligopurine/oligopyrimidine stretch, or 'GAGA' box, and a downstream 56-bp GAGA box which contains a 10-bp mirror repeat and is sensitive to cleavage with S1 nuclease. Images Figure 1 Figure 3 Figure 4 Figure 6 PMID:7646438

  9. COL1A1 mutation analysis in Lithuanian patients with osteogenesis imperfecta.

    PubMed

    Benusiené, Egle; Kucinskas, Vaidutis

    2003-01-01

    Osteogenesis imperfecta (OI) is a generalised disorder of connective tissue characterised by an increased fragility of bones and also manifested in other tissues containing collagen type I, by blue sclera, hearing loss, dentinogenesis imperfecta, hyperextensible joints, hernias and easy bruising. OI is dominantly inherited and results in >90% OI cases, caused by mutations in one of the two genes COL1A1 or COL1A2 coding for type I procollagen. The Lithuanian OI database comprises 147 case records covering the period of 1980 - 2001. Clinical and genealogical analysis of OI cases/families from Lithuania available for examination revealed 18 familial cases of OI type I and 22 sporadic cases: OI type II (3 cases), OI type III (11 cases) and OI type I (8 cases). As a result of their molecular genetic investigation, 11 mutations were identified in the COL1A1 gene in 13 unrelated patients. Of them, nine mutations (E500X, G481A, c.2046insCTCTCTAG, c.1668delT, c.1667insC, c.4337insC, IVS19+1G > A, IVS20-2A > G, IVS22-1G > T) appeared to be novel, i.e. not yet registered in the Human Type I and Type III Collagen Mutations Database (http://www.le.ac.uk/genetics/collagen). PMID:12590186

  10. Antisense-induced messenger depletion corrects a COL6A2 dominant mutation in Ullrich myopathy.

    PubMed

    Gualandi, Francesca; Manzati, Elisa; Sabatelli, Patrizia; Passarelli, Chiara; Bovolenta, Matteo; Pellegrini, Camilla; Perrone, Daniela; Squarzoni, Stefano; Pegoraro, Elena; Bonaldo, Paolo; Ferlini, Alessandra

    2012-12-01

    Collagen VI gene mutations cause Ullrich and Bethlem muscular dystrophies. Pathogenic mutations frequently have a dominant negative effect, with defects in collagen VI chain secretion and assembly. It is agreed that, conversely, collagen VI haploinsufficiency has no pathological consequences. Thus, RNA-targeting approaches aimed at preferentially inactivating the mutated COL6 messenger may represent a promising therapeutic strategy. By in vitro studies we obtained the preferential depletion of the mutated COL6A2 messenger, by targeting a common single-nucleotide polymorphism (SNP), cistronic with a dominant COL6A2 mutation. We used a 2'-O-methyl phosphorothioate (2'OMePS) antisense oligonucleotide covering the SNP within exon 3, which is out of frame. Exon 3 skipping has the effect of depleting the mutated transcript via RNA nonsense-mediated decay, recovering the correct collagen VI secretion and restoring the ability to form an interconnected microfilament network into the extracellular matrix. This novel RNA modulation approach to correcting dominant mutations may represent a therapeutic strategy potentially applicable to a great variety of mutations and diseases. PMID:22992134

  11. Loss of col8a1a function during zebrafish embryogenesis results in congenital vertebral malformations.

    PubMed

    Gray, Ryan S; Wilm, Thomas P; Smith, Jeff; Bagnat, Michel; Dale, Rodney M; Topczewski, Jacek; Johnson, Stephen L; Solnica-Krezel, Lilianna

    2014-02-01

    Congenital vertebral malformations (CVM) occur in 1 in 1000 live births and in many cases can cause spinal deformities, such as scoliosis, and result in disability and distress of affected individuals. Many severe forms of the disease, such as spondylocostal dystostosis, are recessive monogenic traits affecting somitogenesis, however the etiologies of the majority of CVM cases remain undetermined. Here we demonstrate that morphological defects of the notochord in zebrafish can generate congenital-type spine defects. We characterize three recessive zebrafish leviathan/col8a1a mutant alleles ((m531, vu41, vu105)) that disrupt collagen type VIII alpha1a (col8a1a), and cause folding of the embryonic notochord and consequently adult vertebral column malformations. Furthermore, we provide evidence that a transient loss of col8a1a function or inhibition of Lysyl oxidases with drugs during embryogenesis was sufficient to generate vertebral fusions and scoliosis in the adult spine. Using periodic imaging of individual zebrafish, we correlate focal notochord defects of the embryo with vertebral malformations (VM) in the adult. Finally, we show that bends and kinks in the notochord can lead to aberrant apposition of osteoblasts normally confined to well-segmented areas of the developing vertebral bodies. Our results afford a novel mechanism for the formation of VM, independent of defects of somitogenesis, resulting from aberrant bone deposition at regions of misshapen notochord tissue. PMID:24333517

  12. Variants in a Novel Epidermal Collagen Gene (COL29A1) Are Associated with Atopic Dermatitis

    PubMed Central

    Kerscher, Tamara; Rüschendorf, Franz; Esparza-Gordillo, Jorge; Worm, Margitta; Gruber, Christoph; Mayr, Gabriele; Albrecht, Mario; Rohde, Klaus; Schulz, Herbert; Wahn, Ulrich; Hubner, Norbert; Lee, Young-Ae

    2007-01-01

    Atopic dermatitis (AD) is a common chronic inflammatory skin disorder and a major manifestation of allergic disease. AD typically presents in early childhood often preceding the onset of an allergic airway disease, such as asthma or hay fever. We previously mapped a susceptibility locus for AD on Chromosome 3q21. To identify the underlying disease gene, we used a dense map of microsatellite markers and single nucleotide polymorphisms, and we detected association with AD. In concordance with the linkage results, we found a maternal transmission pattern. Furthermore, we demonstrated that the same families contribute to linkage and association. We replicated the association and the maternal effect in a large independent family cohort. A common haplotype showed strong association with AD (p = 0.000059). The associated region contained a single gene, COL29A1, which encodes a novel epidermal collagen. COL29A1 shows a specific gene expression pattern with the highest transcript levels in skin, lung, and the gastrointestinal tract, which are the major sites of allergic disease manifestation. Lack of COL29A1 expression in the outer epidermis of AD patients points to a role of collagen XXIX in epidermal integrity and function, the breakdown of which is a clinical hallmark of AD. PMID:17850181

  13. Generation of Col2a1-EGFP iPS Cells for Monitoring Chondrogenic Differentiation

    PubMed Central

    Saito, Taku; Yano, Fumiko; Mori, Daisuke; Ohba, Shinsuke; Hojo, Hironori; Otsu, Makoto; Eto, Koji; Nakauchi, Hiromitsu; Tanaka, Sakae; Chung, Ung-il; Kawaguchi, Hiroshi

    2013-01-01

    Induced pluripotent stem cells (iPSC) are a promising cell source for cartilage regenerative medicine; however, the methods for chondrocyte induction from iPSC are currently developing and not yet sufficient for clinical application. Here, we report the establishment of a fluorescent indicator system for monitoring chondrogenic differentiation from iPSC to simplify screening for effective factors that induce chondrocytes from iPSC. We generated iPSC from embryonic fibroblasts of Col2a1-EGFP transgenic mice by retroviral transduction of Oct4, Sox2, Klf4, and c-Myc. Among the 30 clones of Col2a1-EGFP iPSC we established, two clones showed high expression levels of embryonic stem cell (ESC) marker genes, similar to control ESC. A teratoma formation assay showed that the two clones were pluripotent and differentiated into cell types from all three germ layers. The fluorescent signal was observed during chondrogenic differentiation of the two clones concomitant with the increase in chondrocyte marker expression. In conclusion, Col2a1-EGFP iPSC are useful for monitoring chondrogenic differentiation and will contribute to research in cartilage regenerative medicine. PMID:24066106

  14. Generation of Col2a1-EGFP iPS cells for monitoring chondrogenic differentiation.

    PubMed

    Saito, Taku; Yano, Fumiko; Mori, Daisuke; Ohba, Shinsuke; Hojo, Hironori; Otsu, Makoto; Eto, Koji; Nakauchi, Hiromitsu; Tanaka, Sakae; Chung, Ung-il; Kawaguchi, Hiroshi

    2013-01-01

    Induced pluripotent stem cells (iPSC) are a promising cell source for cartilage regenerative medicine; however, the methods for chondrocyte induction from iPSC are currently developing and not yet sufficient for clinical application. Here, we report the establishment of a fluorescent indicator system for monitoring chondrogenic differentiation from iPSC to simplify screening for effective factors that induce chondrocytes from iPSC. We generated iPSC from embryonic fibroblasts of Col2a1-EGFP transgenic mice by retroviral transduction of Oct4, Sox2, Klf4, and c-Myc. Among the 30 clones of Col2a1-EGFP iPSC we established, two clones showed high expression levels of embryonic stem cell (ESC) marker genes, similar to control ESC. A teratoma formation assay showed that the two clones were pluripotent and differentiated into cell types from all three germ layers. The fluorescent signal was observed during chondrogenic differentiation of the two clones concomitant with the increase in chondrocyte marker expression. In conclusion, Col2a1-EGFP iPSC are useful for monitoring chondrogenic differentiation and will contribute to research in cartilage regenerative medicine. PMID:24066106

  15. Hair follicle aging is driven by transepidermal elimination of stem cells via COL17A1 proteolysis.

    PubMed

    Matsumura, Hiroyuki; Mohri, Yasuaki; Binh, Nguyen Thanh; Morinaga, Hironobu; Fukuda, Makoto; Ito, Mayumi; Kurata, Sotaro; Hoeijmakers, Jan; Nishimura, Emi K

    2016-02-01

    Hair thinning and loss are prominent aging phenotypes but have an unknown mechanism. We show that hair follicle stem cell (HFSC) aging causes the stepwise miniaturization of hair follicles and eventual hair loss in wild-type mice and in humans. In vivo fate analysis of HFSCs revealed that the DNA damage response in HFSCs causes proteolysis of type XVII collagen (COL17A1/BP180), a critical molecule for HFSC maintenance, to trigger HFSC aging, characterized by the loss of stemness signatures and by epidermal commitment. Aged HFSCs are cyclically eliminated from the skin through terminal epidermal differentiation, thereby causing hair follicle miniaturization. The aging process can be recapitulated by Col17a1 deficiency and prevented by the forced maintenance of COL17A1 in HFSCs, demonstrating that COL17A1 in HFSCs orchestrates the stem cell-centric aging program of the epithelial mini-organ. PMID:26912707

  16. The ColRS system of Xanthomonas oryzae pv. oryzae is required for virulence and growth in iron-limiting conditions.

    PubMed

    Subramoni, Sujatha; Pandey, Alok; Vishnu Priya, M R; Patel, Hitendra Kumar; Sonti, Ramesh V

    2012-09-01

    Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight of rice, produces siderophores only under iron-limiting conditions. We screened 15 400 mTn5-induced mutants of X. oryzae pv. oryzae and isolated 27 mutants that produced siderophores even under iron-replete conditions. We found that the mTn5 insertions in 25 of these mutants were in or close to six genes. Mutants with insertions in five of these genes [colS, XOO1806 (a conserved hypothetical protein), acnB, prpR and prpB] exhibited a deficiency for growth on iron-limiting medium and a decrease in virulence. Insertions in a sixth gene, XOO0007 (a conserved hypothetical protein), were found to affect the ability to grow on iron-limiting medium, but did not affect the virulence. Targeted gene disruptants for colR (encoding the predicted cognate regulatory protein for ColS) also exhibited a deficiency for growth on iron-limiting medium and a decrease in virulence. colR and colS mutants were defective in the elicitation of hypersensitive response symptoms on the nonhost tomato. In addition, colR and colS mutants induced a rice basal defence response, suggesting that they are compromised in the suppression of host innate immunity. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis demonstrated that a functional ColRS system is required for the optimal expression of several genes encoding components of the type 3 secretion system (T3SS) of X. oryzae pv. oryzae. Our results demonstrate the role of several novel genes, including colR/colS, in the promotion of growth on iron-limiting medium and the virulence of X. oryzae pv. oryzae. PMID:22257308

  17. Strain-Dependent Anterior Segment Dysgenesis and Progression to Glaucoma in Col4a1 Mutant Mice

    PubMed Central

    Mao, Mao; Smith, Richard S.; Alavi, Marcel V.; Marchant, Jeffrey K.; Cosma, Mihai; Libby, Richard T.; John, Simon W. M.; Gould, Douglas B.

    2015-01-01

    Purpose Mutations in the gene encoding collagen type IV alpha 1 (COL4A1) cause multisystem disorders including anterior segment dysgenesis (ASD) and optic nerve hypoplasia. The penetrance and severity of individual phenotypes depends on genetic context. Here, we tested the effects of a Col4a1 mutation in two different genetic backgrounds to compare how genetic context influences ocular dysgenesis, IOP, and progression to glaucoma. Methods Col4a1 mutant mice maintained on a C57BL/6J background were crossed to either 129S6/SvEvTac or CAST/EiJ and the F1 progeny were analyzed by slit-lamp biomicroscopy and optical coherence tomography. We also measured IOPs and compared tissue sections of eyes and optic nerves. Results. We found that the CAST/EiJ inbred strain has a relatively uniform and profound suppression on the effects of Col4a1 mutation and that mutant CASTB6F1 mice were generally only very mildly affected. In contrast, mutant 129B6F1 mice had more variable and severe ASD and IOP dysregulation that were associated with glaucomatous signs including lost or damaged retinal ganglion cell axons and excavation of the optic nerve head. Conclusions. Ocular defects in Col4a1 mutant mice model ASD and glaucoma that are observed in a subset of patients with COL4A1 mutations. We demonstrate that different inbred strains of mice give graded severities of ASD and we detected elevated IOP and glaucomatous damage in 129B6F1, but not CASTB6F1 mice that carried a Col4a1 mutation. These data demonstrate that genetic context differences are one factor that may contribute to the variable penetrance and severity of ASD and glaucoma in patients with COL4A1 mutations. PMID:26567795

  18. Dominant and Recessive Forms of Fibrochondrogenesis Resulting from Mutations at a Second Locus, COL11A2

    PubMed Central

    Tompson, Stuart W.; Faqeih, Eissa Ali; Ala-Kokko, Leena; Hecht, Jacqueline T.; Miki, Rika; Funari, Tara; Funari, Vincent A.; Nevarez, Lisette; Krakow, Deborah; Cohn, Daniel H.

    2011-01-01

    Fibrochondrogenesis is a severe, recessively inherited skeletal dysplasia shown to result from mutations in the gene encoding the proα1(XI) chain of type XI collagen, COL11A1. The first of two cases reported here was the affected offspring of first cousins and sequence analysis excluded mutations in COL11A1. Consequently, whole-genome SNP genotyping was performed to identify blocks of homozygosity, identical-by-descent, wherein the disease locus would reside. COL11A1 was not within a region of homozygosity, further excluding it as the disease locus, but the gene encoding the proα2(XI) chain of type XI collagen, COL11A2, was located within a large region of homozygosity. Sequence analysis identified homozygosity for a splice donor mutation in intron 18. Exon trapping demonstrated that the mutation resulted in skipping of exon 18 and predicted deletion of 18 amino acids from the triple helical domain of the protein. In the second case, heterozygosity for a de novo 9 bp deletion in exon 40 of COL11A2 was identified, indicating that there are autosomal dominant forms of fibrochondrogenesis. These findings thus demonstrate that fibrochondrogenesis can result from either recessively- or dominantly-inherited mutations in COL11A2. PMID:22246659

  19. Genetic association of COL1A1 polymorphisms with high myopia in Asian population: a Meta-analysis

    PubMed Central

    Gong, Bo; Qu, Chao; Huang, Xiao-Fang; Ye, Zi-Meng; Zhang, Ding-Ding; Shi, Yi; Chen, Rong; Liu, Yu-Ping; Shuai, Ping

    2016-01-01

    AIM To comprehensively evaluate the potential association of COL1A1 polymorphisms with high myopia by a systematic review and Meta-analysis. METHODS All association studies on COL1A1 and high myopia reported up to June 10, 2014 in PubMed, Embase, Web of Science, and the Chinese Biomedical Database were retrieved. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were analyzed for single-nucleotide polymorphisms (SNPs) using fixed- and random- effects models according to between-study heterogeneity. Publication bias analyses were conducted by Egger's test. RESULTS A total of four studies from reported papers were included in this analysis. The Meta-analyses for COL1A1 rs2075555, composed of 2304 high myopia patients and 2272 controls, failed to detect any significant association with high myopia. A total of 971 cases and 649 controls were tested for COL1A1 rs2269336. The association of COL1A1 rs2269336 with high myopia was observed in recessive model (CC vs CG+GG, P=0.03) and in heterozygous model (CG vs GG, P=0.04), but not in other models. CONCLUSION This Meta-analysis shows that COL1A1 rs2269336 (CC vs CG+GG) affects individual susceptibility to high myopia, whereas there is no association detected between SNPs rs2075555 and high myopia. Given the limited sample size, further investigations including more ethnic groups are required to validate the association. PMID:27588274

  20. Ehlers-Danlos syndrome type IV is associated with a novel G984R COL3A1 mutation.

    PubMed

    Deng, Yao; Wei, Shijie; Hu, Shijun; Chen, Jinlan; Tan, Zhiping; Yang, Yifeng

    2015-07-01

    Ehlers-Danlos syndrome type IV is an autosomal dominant connective tissue disease. Mutations in COL3A1 have been identified to underlie this disease; however, to the best of our knowledge, no COL3A1 mutations have been reported in Ehlers-Danlos syndrome type IV patients with an ascending aortic aneurysm. In order to develop further understanding of COL3A1 mutations, an Ehlers-Danlos syndrome type IV patient diagnosed with an ascending aortic aneurysm and a familial history of sudden mortality was analyzed. Genomic DNA was isolated from the peripheral blood of the patient and his family members. All coding exons of eight aneurysm-related genes (FBN1, TGFBR1, TGFBR 2, MYH11, ACTA2, SLC2A10, NOTCH1 and COL3A1) were amplified using polymerase chain reaction (PCR). The PCR products were sequenced with the ABI 3100 Genetic Analyzer, and a mutation was predicted and identified using Polyphen-2, SIFT and Mutation Taster. The novel mutation was identified as c.2950G>A in COL3A1, which results in p.G984R. All three programs predicted this mutation to be deleterous to the protein function. The novel mutation identified in this study is potentially responsible for Ehlers-Danlos syndrome type IV in this patient, and expands the spectrum of COL3A1 mutations. PMID:25776230

  1. Premature termination codons in the Type VII collagen gene (COL7A1) underlie severe, mutilating recessive dystrophic epidermolysis bullosa

    SciTech Connect

    Christiano, A.M.; Uitto, J. ); Anhalt, G. ); Gibbons, S.; Bauer, E.A. )

    1994-05-01

    Epidermolysis bullosa (EB) is a group of heritable mechano-bullous skin diseases classified into three major categories on the basis of the level of tissue separation within the dermal-epidermal basement membrane zone. The most severe, dystrophic (scarring) forms of EB demonstrate blister formation below the cutaneous basement membrane at the level of the anchoring fibrils. Ultrastructural observations of altered anchoring fibrils and genetic linkage to the gene encoding type VII collagen (COL7A1), the major component of anchoring fibrils, have implicated COL7A1 as the candidate gene in the dystrophic forms of EB. The authors have recently cloned the entire cDNA and gene for human COL7A1, which has been mapped to 3p21. In this study, they describe mutations in four COL7A1 alleles in three patients with severe, mutilating recessive dystrophic EB (Hallopeau-Siemens type, HS-RDEB). Each of these mutations resulted in a premature termination codon (PTC) in the amino-terminal portion of COL7A1. One of the patients was a compound heterozygote for two different mutations. The heterozygous carriers showed an [approximately] 50% reduction in anchoring fibrils, yet were clinically unaffected. Premature termination codons in both alleles of COL7A1 may thus be a major underlying cause of the severe, recessive dystrophic forms of EB. 40 refs., 8 figs.

  2. COL11A1 confers chemoresistance on ovarian cancer cells through the activation of Akt/c/EBPβ pathway and PDK1 stabilization

    PubMed Central

    Wu, Yi-Hui; Chang, Tzu-Hao; Huang, Yu-Fang; Chen, Chien-Chin; Chou, Cheng-Yang

    2015-01-01

    Chemoresistance to anticancer drugs substantially reduces survival in epithelial ovarian carcinoma (EOC). Here, microarray analysis showed that collagen type XI alpha 1 (COL11A1) is a chemotherapy response-associated gene. Chemoresistant cells expressed higher COL11A1 and c/EBPβ than chemosensitive cells. COL11A1 or c/EBPβ downregulation suppressed chemoresistance, whereas COL11A1 overexpression attenuated sensitivity to cisplatin and paclitaxel. The c/EBPβ binding site in the COL11A1 promoter was identified as the major determinant of cisplatin- and paclitaxel-induced COL11A1 expression. Immunoprecipitation and immunofluorescence showed that in resistant cells, Akt and PDK1 were highly expressed and that anticancer drugs enhanced binding activity between COL11A1 and PDK1 binding and attenuated PDK1 ubiquitination and degradation. Conversely, chemosensitive cells showed decreased activity of COL11A1 binding to PDK1 and increased PDK1 ubiquitination, which were reversed by COL11A1 overexpression. Analysis of 104 EOC patients showed that high COL11A1 mRNA levels are significantly associated with poor chemoresponse and clinical outcome. PMID:26087191

  3. Homozygosity and Heterozygosity for Null Col5a2 Alleles Produce Embryonic Lethality and a Novel Classic Ehlers-Danlos Syndrome-Related Phenotype.

    PubMed

    Park, Arick C; Phillips, Charlotte L; Pfeiffer, Ferris M; Roenneburg, Drew A; Kernien, John F; Adams, Sheila M; Davidson, Jeffrey M; Birk, David E; Greenspan, Daniel S

    2015-07-01

    Null alleles for the COL5A1 gene and missense mutations for COL5A1 or the COL5A2 gene underlie cases of classic Ehlers-Danlos syndrome, characterized by fragile, hyperextensible skin and hypermobile joints. However, no classic Ehlers-Danlos syndrome case has yet been associated with COL5A2 null alleles, and phenotypes that might result from such alleles are unknown. We describe mice with null alleles for the Col5a2. Col5a2(-/-) homozygosity is embryonic lethal at approximately 12 days post conception. Unlike previously described mice null for Col5a1, which die at 10.5 days post conception and virtually lack collagen fibrils, Col5a2(-/-) embryos have readily detectable collagen fibrils, thicker than in wild-type controls. Differences in Col5a2(-/-) and Col5a1(-/-) fibril formation and embryonic survival suggest that α1(V)3 homotrimers, a rare collagen V isoform that occurs in the absence of sufficient levels of α2(V) chains, serve functional roles that partially compensate for loss of the most common collagen V isoform. Col5a2(+/-) adults have skin with marked hyperextensibility and reduced tensile strength at high strain but not at low strain. Col5a2(+/-) adults also have aortas with increased compliance and reduced tensile strength. Results thus suggest that COL5A2(+/-) humans, although unlikely to present with frank classic Ehlers-Danlos syndrome, are likely to have fragile connective tissues with increased susceptibility to trauma and certain chronic pathologic conditions. PMID:25987251

  4. Insulation of the ubiquitous Rxrb promoter from the cartilage-specific adjacent gene, Col11a2.

    PubMed

    Murai, Junko; Ikegami, Daisuke; Okamoto, Mina; Yoshikawa, Hideki; Tsumaki, Noriyuki

    2008-10-10

    The retinoid X receptor beta gene (Rxrb) is located just upstream of the alpha2(XI) collagen chain gene (Col11a2) in a head-to-tail manner. However, the domain structures of these genes are unknown. Col11a2 is specifically expressed in cartilage. In the present study, we found Rxrb expression in various tissues with low expression in the cartilage. Col11a2 1st intron enhancer directed cartilage specific expression when linked to the heterologous promoter in transgenic mice. These results suggest the presence of enhancer-blocking elements that insulate Rxrb promoter from the Col11a2 enhancer. So far, most of insulators examined in vertebrates contain a binding site for CTCF. We found two possible CTCF-binding sites: one (11P) in the intergenic region between Rxrb and Col11a2 by electrophoretic mobility shift assays, and the other in the 4th intron of RXRB by data base search. To examine the function of these elements, we prepared bacterial artificial chromosome (BAC) transgene constructs containing a 142-kb genomic DNA insert with RXRB and COL11A2 sequences in the middle. Mutation of 11P significantly decreased the RXRB promoter activity in muscular cells and significantly increased expression levels of RXRB in chondrosarcoma cells. In transgenic mouse assays, the wild-type BAC transgene partly recapitulated endogenous Rxrb expression patterns. A 507-bp deletion mutation including 11P enhanced the cartilage-specific activity of the RXRB promoter in BAC transgenic mice. Chromatin immunoprecipitation analysis showed that CTCF was associated with RX4, but not with 11P. Our results showed that the intergenic sequence including 11P insulates Rxrb promoter from Col11a2 enhancer, possibly associating with unknown factors that recognize a motif similar to CTCF. PMID:18682388

  5. Stickler syndrome caused by COL2A1 mutations: genotype–phenotype correlation in a series of 100 patients

    PubMed Central

    Hoornaert, Kristien P; Vereecke, Inge; Dewinter, Chantal; Rosenberg, Thomas; Beemer, Frits A; Leroy, Jules G; Bendix, Laila; Björck, Erik; Bonduelle, Maryse; Boute, Odile; Cormier-Daire, Valerie; De Die-Smulders, Christine; Dieux-Coeslier, Anne; Dollfus, Hélène; Elting, Mariet; Green, Andrew; Guerci, Veronica I; Hennekam, Raoul C M; Hilhorts-Hofstee, Yvonne; Holder, Muriel; Hoyng, Carel; Jones, Kristi J; Josifova, Dragana; Kaitila, Ilkka; Kjaergaard, Suzanne; Kroes, Yolande H; Lagerstedt, Kristina; Lees, Melissa; LeMerrer, Martine; Magnani, Cinzia; Marcelis, Carlo; Martorell, Loreto; Mathieu, Michèle; McEntagart, Meriel; Mendicino, Angela; Morton, Jenny; Orazio, Gabrielli; Paquis, Véronique; Reish, Orit; Simola, Kalle O J; Smithson, Sarah F; Temple, Karen I; Van Aken, Elisabeth; Van Bever, Yolande; van den Ende, Jenneke; Van Hagen, Johanna M; Zelante, Leopoldo; Zordania, Riina; De Paepe, Anne; Leroy, Bart P; De Buyzere, Marc; Coucke, Paul J; Mortier, Geert R

    2010-01-01

    Stickler syndrome is an autosomal dominant connective tissue disorder caused by mutations in different collagen genes. The aim of our study was to define more precisely the phenotype and genotype of Stickler syndrome type 1 by investigating a large series of patients with a heterozygous mutation in COL2A1. In 188 probands with the clinical diagnosis of Stickler syndrome, the COL2A1 gene was analyzed by either a mutation scanning technique or bidirectional fluorescent DNA sequencing. The effect of splice site alterations was investigated by analyzing mRNA. Multiplex ligation-dependent amplification analysis was used for the detection of intragenic deletions. We identified 77 different COL2A1 mutations in 100 affected individuals. Analysis of the splice site mutations showed unusual RNA isoforms, most of which contained a premature stop codon. Vitreous anomalies and retinal detachments were found more frequently in patients with a COL2A1 mutation compared with the mutation-negative group (P<0.01). Overall, 20 of 23 sporadic patients with a COL2A1 mutation had either a cleft palate or retinal detachment with vitreous anomalies. The presence of vitreous anomalies, retinal tears or detachments, cleft palate and a positive family history were shown to be good indicators for a COL2A1 defect. In conclusion, we confirm that Stickler syndrome type 1 is predominantly caused by loss-of-function mutations in the COL2A1 gene as >90% of the mutations were predicted to result in nonsense-mediated decay. On the basis of binary regression analysis, we developed a scoring system that may be useful when evaluating patients with Stickler syndrome. PMID:20179744

  6. Association of COL1A1 polymorphisms with osteoporosis: a meta-analysis of clinical studies

    PubMed Central

    Xie, Peigen; Liu, Bin; Zhang, Liangming; Chen, Ruiqiang; Yang, Bu; Dong, Jianwen; Rong, Limin

    2015-01-01

    Objective: To conduct a meta-analysis of all association studies on two of the collagen 1 alpha 1 (COL1A1) gene polymorphisms, the -1997G/T (rs1107946) and the -1663indelT (rs2412298) polymorphisms and osteoporosis/BMD and fracture. Methods: PubMed/Medline and Web of Knowledge were searched for relevant association studies published in English. Pooled OR and its corresponding 95% CI or pooled MD and its corresponding 95% CI was calculated with the Cochrane Review Manager (Revman, version 5.2) using a random-effect or a fixed effect model. Results: No significant association between the -1997G/T polymorphism and Lumbar Spine (LS) and Femoral Neck (FN) BMD except for the Caucasian subpopulation wherein subjects with the T allele of the -1997G/T polymorphism was associated with significantly higher LS BMD. Our analysis did reveal that women, especially postmenopausal or perimenopausal women with the GG genotype, had significantly higher Total Hip (TH) BMD than those with the GT. Additionally, our meta-analysis did not show significant association between the -1997G/T polymorphism and risk of fracture, between the -1663indelT polymorphism and LS BMD in postmenopausal or perimenopausal women, or between the -1663indelT polymorphism and the risk of fracture. Conclusions: Our results suggested the possibility of the COL1A1 -1997G/T and the -1663indelT polymorphisms individually playing very little role in osteoporosis and fracture, although more studies are needed especially for the analysis of association between these two polymorphisms and fracture. Haplotype studies may become one important future direction of study to further elucidate whether and how various COL1A1 polymorphisms affect bone health, osteoporosis and fracture. PMID:26628959

  7. A novel COL11A1 mutation affecting splicing in a patient with Stickler syndrome

    PubMed Central

    Kohmoto, Tomohiro; Naruto, Takuya; Kobayashi, Haruka; Watanabe, Miki; Okamoto, Nana; Masuda, Kiyoshi; Imoto, Issei; Okamoto, Nobuhiko

    2015-01-01

    Stickler syndrome is a clinically and genetically heterogeneous collagenopathy characterized by ocular, auditory, skeletal and orofacial abnormalities, commonly occurring as an autosomal dominant trait. We conducted target resequencing to analyze candidate genes associated with known clinical phenotypes from a 4-year-old girl with Stickler syndrome. We detected a novel heterozygous intronic mutation (NM_001854.3:c.3168+5G>A) in COL11A1 that may impair splicing, which was suggested by in silico prediction and a minigene assay. PMID:27081549

  8. The ColRS system is essential for the hunger response of glucose-growing Pseudomonas putida

    PubMed Central

    2011-01-01

    Background The survival of bacteria largely depends on signaling systems that coordinate cell responses to environmental cues. Previous studies on the two-component ColRS signal system in Pseudomonas putida revealed a peculiar subpopulation lysis phenotype of colR mutant that grows on solid glucose medium. Here, we aimed to clarify the reasons for the lysis of bacteria. Results We present evidence that the lysis defect of P. putida colR mutant is linked to hunger response. A subpopulation prone to lysis was located in the periphery of bacterial cultures growing on solid medium. Cell lysis was observed in glucose-limiting, but not in glucose-rich conditions. Furthermore, lysis was also alleviated by exhaustion of glucose from the medium which was evidenced by a lower lysis of central cells compared to peripheral ones. Thus, lysis takes place at a certain glucose concentration range that most probably provides bacteria a hunger signal. An analysis of membrane protein pattern revealed several hunger-induced changes in the bacterial outer membrane: at glucose limitation the amount of OprB1 channel protein was significantly increased whereas that of OprE was decreased. Hunger-induced up-regulation of OprB1 correlated in space and time with the lysis of the colR mutant, indicating that hunger response is detrimental to the colR-deficient bacteria. The amount of OprB1 is controlled post-transcriptionally and derepression of OprB1 in glucose-limiting medium depends at least partly on the carbon catabolite regulator protein Crc. The essentiality of ColR in hunger response can be bypassed by reducing the amount of certain outer membrane proteins. In addition to depletion of OprB1, the lysis defect of colR mutant can be suppressed by the down-regulation of OprF levels and the hindering of SecB-dependent protein secretion. Conclusions We show that Pseudomonas putida growing on solid glucose medium adapts to glucose limitation through up-regulation of the sugar channel protein

  9. A novel mutation in COL4A1 gene: a possible cause of early postnatal cerebrovascular events.

    PubMed

    Decio, Alice; Tonduti, Davide; Pichiecchio, Anna; Vetro, Annalisa; Ciccone, Roberto; Limongelli, Ivan; Giorda, Roberto; Caffi, Lorella; Balottin, Umberto; Zuffardi, Orsetta; Orcesi, Simona

    2015-04-01

    COL4A1 is located in humans on chromosome13q34 and it encodes the alpha 1 chain of type IV collagen, a component of basal membrane. It is expressed mainly in the brain, muscles, kidneys and eyes. Different COL4A1 mutations have been reported in many patients who present a very wide spectrum of clinical symptoms. They typically show a multisystemic phenotype. Here we report on the case of a patient carrying a novel de novo splicing mutation of COL4A1 associated with a distinctive clinical picture characterized by onset in infancy and an unusual evolution of the neuroradiological features. At three months of age, the child was diagnosed with a congenital cataract, while his brain MRI was normal. Over the following years, the patient developed focal epilepsy, mild diplegia, asymptomatic microhematuria, raised creatine kinase levels, MRI white matter abnormalities and brain calcification on CT. During the neuroradiological follow-up the extension and intensity of the brain lesions progressively decreased. The significance of a second variant in COL4A1 carried by the child and inherited from his father remains to be clarified. In conclusion, our patient shows new aspects of this collagenopathy and possibly a COL4A1 compound heterozygosity. PMID:25706114

  10. A Gene Gun-mediated Nonviral RNA trans-splicing Strategy for Col7a1 Repair.

    PubMed

    Peking, Patricia; Koller, Ulrich; Hainzl, Stefan; Kitzmueller, Sophie; Kocher, Thomas; Mayr, Elisabeth; Nyström, Alexander; Lener, Thomas; Reichelt, Julia; Bauer, Johann W; Murauer, Eva M

    2016-01-01

    RNA trans-splicing represents an auspicious option for the correction of genetic mutations at RNA level. Mutations within COL7A1 causing strong reduction or absence of type VII collagen are associated with the severe skin blistering disease dystrophic epidermolysis bullosa. The human COL7A1 mRNA constitutes a suitable target for this RNA therapy approach, as only a portion of the almost 9 kb transcript has to be delivered into the target cells. Here, we have proven the feasibility of 5' trans-splicing into the Col7a1 mRNA in vitro and in vivo. We designed a 5' RNA trans-splicing molecule, capable of replacing Col7a1 exons 1-15 and verified it in a fluorescence-based trans-splicing model system. Specific and efficient Col7a1 trans-splicing was confirmed in murine keratinocytes. To analyze trans-splicing in vivo, we used gene gun delivery of a minicircle expressing a FLAG-tagged 5' RNA trans-splicing molecule into the skin of wild-type mice. Histological and immunofluorescence analysis of bombarded skin sections revealed vector delivery and expression within dermis and epidermis. Furthermore, we have detected trans-spliced type VII collagen protein using FLAG-tag antibodies. In conclusion, we describe a novel in vivo nonviral RNA therapy approach to restore type VII collagen expression for causative treatment of dystrophic epidermolysis bullosa. PMID:26928235

  11. A founder mutation in COL4A3 causes autosomal recessive Alport syndrome in the Ashkenazi Jewish population.

    PubMed

    Webb, B D; Brandt, T; Liu, L; Jalas, C; Liao, J; Fedick, A; Linderman, M D; Diaz, G A; Kornreich, R; Trachtman, H; Mehta, L; Edelmann, L

    2014-08-01

    Alport syndrome is an inherited progressive nephropathy arising from mutations in the type IV collagen genes, COL4A3, COL4A4, and COL4A5. Symptoms also include sensorineural hearing loss and ocular lesions. We determined the molecular basis of Alport syndrome in a non-consanguineous Ashkenazi Jewish family with multiple affected females using linkage analysis and next generation sequencing. We identified a homozygous COL4A3 mutation, c.40_63del, in affected individuals with mutant alleles inherited from each parent on partially conserved haplotypes. Large-scale population screening of 2017 unrelated Ashkenazi Jewish samples revealed a carrier frequency of 1 in 183 indicating that COL4A3 c.40_63del is a founder mutation which may be a common cause of Alport syndrome in this population. Additionally, we determined that heterozygous mutation carriers in this family do not meet criteria for a diagnosis of Thin Basement Membrane Nephropathy and concluded that carriers of c.40_63del are not likely to develop benign familial hematuria. PMID:23927549

  12. Mutations in collagen, type XVII, alpha 1 (COL17A1) cause epithelial recurrent erosion dystrophy (ERED).

    PubMed

    Jonsson, Frida; Byström, Berit; Davidson, Alice E; Backman, Ludvig J; Kellgren, Therese G; Tuft, Stephen J; Koskela, Timo; Rydén, Patrik; Sandgren, Ola; Danielson, Patrik; Hardcastle, Alison J; Golovleva, Irina

    2015-04-01

    Corneal dystrophies are a clinically and genetically heterogeneous group of inherited disorders that bilaterally affect corneal transparency. They are defined according to the corneal layer affected and by their genetic cause. In this study, we identified a dominantly inherited epithelial recurrent erosion dystrophy (ERED)-like disease that is common in northern Sweden. Whole-exome sequencing resulted in the identification of a novel mutation, c.2816C>T, p.T939I, in the COL17A1 gene, which encodes collagen type XVII alpha 1. The variant segregated with disease in a genealogically expanded pedigree dating back 200 years. We also investigated a unique COL17A1 synonymous variant, c.3156C>T, identified in a previously reported unrelated dominant ERED-like family linked to a locus on chromosome 10q23-q24 encompassing COL17A1. We show that this variant introduces a cryptic donor site resulting in aberrant pre-mRNA splicing and is highly likely to be pathogenic. Bi-allelic COL17A1 mutations have previously been associated with a recessive skin disorder, junctional epidermolysis bullosa, with recurrent corneal erosions being reported in some cases. Our findings implicate presumed gain-of-function COL17A1 mutations causing dominantly inherited ERED and improve understanding of the underlying pathology. PMID:25676728

  13. COL1A1 transgene expression in stably transfected osteoblastic cells. Relative contributions of first intron, 3'-flanking sequences, and sequences derived from the body of the human COL1A1 minigene

    NASA Technical Reports Server (NTRS)

    Breault, D. T.; Lichtler, A. C.; Rowe, D. W.

    1997-01-01

    Collagen reporter gene constructs have be used to identify cell-specific sequences needed for transcriptional activation. The elements required for endogenous levels of COL1A1 expression, however, have not been elucidated. The human COL1A1 minigene is expressed at high levels and likely harbors sequence elements required for endogenous levels of activity. Using stably transfected osteoblastic Py1a cells, we studied a series of constructs (pOBColCAT) designed to characterize further the elements required for high level of expression. pOBColCAT, which contains the COL1A1 first intron, was expressed at 50-100-fold higher levels than ColCAT 3.6, which lacks the first intron. This difference is best explained by improved mRNA processing rather than a transcriptional effect. Furthermore, variation in activity observed with the intron deletion constructs is best explained by altered mRNA splicing. Two major regions of the human COL1A1 minigene, the 3'-flanking sequences and the minigene body, were introduced into pOBColCAT to assess both transcriptional enhancing activity and the effect on mRNA stability. Analysis of the minigene body, which includes the first five exons and introns fused with the terminal six introns and exons, revealed an orientation-independent 5-fold increase in CAT activity. In contrast the 3'-flanking sequences gave rise to a modest 61% increase in CAT activity. Neither region increased the mRNA half-life of the parent construct, suggesting that CAT-specific mRNA instability elements may serve as dominant negative regulators of stability. This study suggests that other sites within the body of the COL1A1 minigene are important for high expression, e.g. during periods of rapid extracellular matrix production.

  14. COL4A6 is dispensable for autosomal recessive Alport syndrome

    PubMed Central

    Murata, Tomohiro; Katayama, Kan; Oohashi, Toshitaka; Jahnukainen, Timo; Yonezawa, Tomoko; Sado, Yoshikazu; Ishikawa, Eiji; Nomura, Shinsuke; Tryggvason, Karl; Ito, Masaaki

    2016-01-01

    Alport syndrome is caused by mutations in the genes encoding α3, α4, or α5 (IV) chains. Unlike X-linked Alport mice, α5 and α6 (IV) chains are detected in the glomerular basement membrane of autosomal recessive Alport mice, however, the significance of this finding remains to be investigated. We therefore generated mice lacking both α3 and α6 (IV) chains and compared their renal function and survival with Col4a3 knockout mice of 129 × 1/Sv background. No significant difference was observed in the renal function or survival of the two groups, or when the mice were backcrossed once to C57BL/6 background. However, the survival of backcrossed double knockout mice was significantly longer than that of the mice of 129 × 1/Sv background, which suggests that other modifier genes were involved in this phenomenon. In further studies we identified two Alport patients who had a homozygous mutation in intron 46 of COL4A4. The α5 and α6 (IV) chains were focally detected in the glomerular basement membrane of these patients. These findings indicate that although α5 and α6 (IV) chains are induced in the glomerular basement membrane in autosomal recessive Alport syndrome, their induction does not seem to play a major compensatory role. PMID:27377778

  15. Artificial Autopolyploidization Modifies the Tricarboxylic Acid Cycle and GABA Shunt in Arabidopsis thaliana Col-0

    NASA Astrophysics Data System (ADS)

    Vergara, Fredd; Kikuchi, Jun; Breuer, Christian

    2016-05-01

    Autopolyploidy is a process whereby the chromosome set is multiplied and it is a common phenomenon in angiosperms. Autopolyploidy is thought to be an important evolutionary force that has led to the formation of new plant species. Despite its relevance, the consequences of autopolyploidy in plant metabolism are poorly understood. This study compares the metabolic profiles of natural diploids and artificial autotetraploids of Arabidopsis thaliana Col-0. Different physiological parameters are compared between diploids and autotetraploids using nuclear magnetic resonance (NMR), elemental analysis (carbon:nitrogen balance) and quantitative real-time PCR (qRT-PCR). The main difference between diploid and autotetraploid A. thaliana Col-0 is observed in the concentration of metabolites related to the tricarboxylic acid cycle (TCA) and γ-amino butyric acid (GABA) shunt, as shown by multivariate statistical analysis of NMR spectra. qRT-PCR shows that genes related to the TCA and GABA shunt are also differentially expressed between diploids and autotetraploids following similar trends as their corresponding metabolites. Solid evidence is presented to demonstrate that autopolyploidy influences core plant metabolic processes.

  16. Artificial Autopolyploidization Modifies the Tricarboxylic Acid Cycle and GABA Shunt in Arabidopsis thaliana Col-0

    PubMed Central

    Vergara, Fredd; Kikuchi, Jun; Breuer, Christian

    2016-01-01

    Autopolyploidy is a process whereby the chromosome set is multiplied and it is a common phenomenon in angiosperms. Autopolyploidy is thought to be an important evolutionary force that has led to the formation of new plant species. Despite its relevance, the consequences of autopolyploidy in plant metabolism are poorly understood. This study compares the metabolic profiles of natural diploids and artificial autotetraploids of Arabidopsis thaliana Col-0. Different physiological parameters are compared between diploids and autotetraploids using nuclear magnetic resonance (NMR), elemental analysis (carbon:nitrogen balance) and quantitative real-time PCR (qRT-PCR). The main difference between diploid and autotetraploid A. thaliana Col-0 is observed in the concentration of metabolites related to the tricarboxylic acid cycle (TCA) and γ-amino butyric acid (GABA) shunt, as shown by multivariate statistical analysis of NMR spectra. qRT-PCR shows that genes related to the TCA and GABA shunt are also differentially expressed between diploids and autotetraploids following similar trends as their corresponding metabolites. Solid evidence is presented to demonstrate that autopolyploidy influences core plant metabolic processes. PMID:27212081

  17. Artificial Autopolyploidization Modifies the Tricarboxylic Acid Cycle and GABA Shunt in Arabidopsis thaliana Col-0.

    PubMed

    Vergara, Fredd; Kikuchi, Jun; Breuer, Christian

    2016-01-01

    Autopolyploidy is a process whereby the chromosome set is multiplied and it is a common phenomenon in angiosperms. Autopolyploidy is thought to be an important evolutionary force that has led to the formation of new plant species. Despite its relevance, the consequences of autopolyploidy in plant metabolism are poorly understood. This study compares the metabolic profiles of natural diploids and artificial autotetraploids of Arabidopsis thaliana Col-0. Different physiological parameters are compared between diploids and autotetraploids using nuclear magnetic resonance (NMR), elemental analysis (carbon:nitrogen balance) and quantitative real-time PCR (qRT-PCR). The main difference between diploid and autotetraploid A. thaliana Col-0 is observed in the concentration of metabolites related to the tricarboxylic acid cycle (TCA) and γ-amino butyric acid (GABA) shunt, as shown by multivariate statistical analysis of NMR spectra. qRT-PCR shows that genes related to the TCA and GABA shunt are also differentially expressed between diploids and autotetraploids following similar trends as their corresponding metabolites. Solid evidence is presented to demonstrate that autopolyploidy influences core plant metabolic processes. PMID:27212081

  18. Purification and characterization of a novel collagenase from Bacillus pumilus Col-J.

    PubMed

    Wu, Qi; Li, Chen; Li, Chenglei; Chen, Hui; Shuliang, Liu

    2010-01-01

    The collagenase, produced extracellular by Bacillus pumilus Col-J, was purified by ammonium sulfate precipitation followed by two gel filtrations, involving Sephadex G-100 column and Sepharose Fast Flow column. Purified collagenase has a 31.53-fold increase in specific activity of 87.33 U/mg and 7.00% recovery. The collagenase has a relative molecular weight of 58.64 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal temperature for the enzyme reaction was 45 degrees C. More than 50% of the original activity still remained after 5 min of incubation at 70 degrees C or 10 min at 60 degrees C. The maximal enzyme activity of collagenase was obtained at pH 7.5, and it was stable over a pH range of 6.5-8.0. The collagenase activity was strongly inhibited by Mn(2+), Pb(2+), ethylenediamine tetraacetic acid, ethylene glycol tetraacetic acid, and beta-mercaptoethanol. However, Ca(2+) and Mg(2+) greatly increased its activity. The collagenase from B. pumilus Col-J showed highly specific activity towards the native collagen from calf skin. The K(m) and V(max) of the enzyme for collagen were 0.79 mg/mL and 129.5 U, respectively. PMID:19475515

  19. Common ancestry of three Ashkenazi-American families with Alport syndrome and COL4A5 R1677Q.

    PubMed

    Barker, D F; Denison, J C; Atkin, C L; Gregory, M C

    1997-05-01

    Mutations in the basement membrane collagen gene COL4A5 cause the progressive renal glomerular nephropathy and typical hearing loss that occur in X-linked Alport syndrome. Nearly all cases involve distinct mutations, as expected for an X-linked disease that significantly reduces the fitness of affected males. A few exceptional COL4A5 mutations appear to be associated with a reduced disease severity and may account for a significant proportion of late-onset Alport syndrome in populations where a founder effect has occurred. The novel mutation reported here, COL4A5 arg1677gln, has been detected in three independently ascertained Ashkenazi-American families, causes a relatively mild form of nephritis with typical onset in the fourth or fifth decade, and may be involved in the etiology of a large proportion of adult-onset hereditary nephritis in Ashkenazi Jews. PMID:9150741

  20. 1,25-Dihydroxyvitamin D3 inhibition of col1a1 promoter expression in calvariae from neonatal transgenic mice

    NASA Technical Reports Server (NTRS)

    Bedalov, A.; Salvatori, R.; Dodig, M.; Kapural, B.; Pavlin, D.; Kream, B. E.; Clark, S. H.; Woody, C. O.; Rowe, D. W.; Lichtler, A. C.

    1998-01-01

    We studied the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on organ cultures of transgenic mouse calvariae containing segments of the Col1a1 promoter extending to -3518, -2297, -1997, -1794, -1763, and -1719 bp upstream of the transcription start site fused to the chloramphenicol acetyltransferase (CAT) reporter gene. 1,25(OH)2D3 had a dose-dependent inhibitory effect on the expression of the -3518 bp promoter construct (ColCAT3.6), with maximal inhibition of about 50% at 10 nM. This level of inhibition was consistent with the previously observed effect on the endogenous Col1a1 gene in bone cell models. All of the shorter constructs were also inhibited by 10 nM 1,25(OH)2D3, suggesting that the sequences required for 1, 25(OH)2D3 inhibition are downstream of -1719 bp. The inhibitory effect of 1,25(OH)2D3 on transgene mRNA was maintained in the presence of the protein synthesis inhibitor cycloheximide, suggesting that the inhibitory effect on Col1a1 gene transcription does not require de novo protein synthesis. We also examined the in vivo effect of 1,25(OH)2D3 treatment of transgenic mice on ColCAT activity, and found that 48 h treatment caused a dose-dependent inhibition of CAT activity in calvariae comparable to that observed in organ cultures. In conclusion, we demonstrated that 1,25(OH)2D3 inhibits Col1A1 promoter activity in transgenic mouse calvariae, both in vivo and in vitro. The results indicate that there is a 1, 25(OH)2D3 responsive element downstream of -1719 bp. The inhibitory effect does not require new protein synthesis.

  1. Structural organization of the human type VII collagen gene (COL7A1), composed of more exons than any previously characterized gene

    SciTech Connect

    Christiano, A.M.; Chung-Honet, L.C.; Greenspan, D.S.; Hoffman, G.G.; Lee, S.; Cheng, W. ); Uitto, J. )

    1994-05-01

    The human type VII collagen (COL7A1) gene is the locus for mutations in at least some cases of dystrophic epidermolysis bullosa. Here the authors describe the entire intron/exon organization of COL7A1, which is shown to have 118 exons, more than any previously described gene. Despite this complexity, COL7A1 is compact. Consisting of 31,132 bp from transcription start site to polyadenylation site, it is only about three times the size of type VII collagen mRNA. Thus, COL7A1 introns are small. A 71-nucleotide COL7A1 intron is the smallest intron yet reported in a collagen gene, and only one COL7A1 intron is greater than 1 kb in length. All exons in the COL7A1 triple helix coding region that do not begin with sequences corresponding to imperfections of the triple helix begin with intact codons for Gly residues of Gly-X-Y repeats. This is reminiscent of the structure of fibrillar rather than other nonfibrillar collagen genes. In addition, the COL7A1 triple helix coding region contains many exons of recurring sizes (e.g., 25 exons are 36 bp, 12 exons are 45 bp, 8 exons are 63 bp), suggesting an evolutionary origin distinct from those of other nonfibrillar collagen genes. Sequences from the 5[prime] portion of COL7A1 are presented along with the 3766-bp intergenic sequence, which separated COL7A1 from the upstream gene encoding the core I protein of the cytochrome bc[sub 1] complex. The COL7A1 promoter region is found to lack extensive homologies with promoter regions of other genes expressed primarily in skin. 60 refs., 5 figs., 1 tab.

  2. COOH-Terminal Collagen Q (COLQ) Mutants Causing Human Deficiency of Endplate Acetylcholinesterase Impair the Interaction of ColQ with Proteins of the Basal Lamina

    PubMed Central

    Arredondo, Juan; Lara, Marian; Ng, Fiona; Gochez, Danielle A.; Lee, Diana C.; Logia, Stephanie P.; Nguyen, Joanna; Maselli, Ricardo A.

    2014-01-01

    Collagen Q (ColQ) is a key multidomain functional protein of the neuromuscular junction (NMJ), crucial for anchoring acetylcholinesterase (AChE) to the basal lamina (BL) and accumulating AChE at the NMJ. The attachment of AChE to the BL is primarily accomplished by the binding of the ColQ collagen domain to the heparan sulfate proteoglycan perlecan and the COOH-terminus to the muscle-specific receptor tyrosine kinase (MuSK), which in turn plays a fundamental role in the development and maintenance of the NMJ. Yet, the precise mechanism by which ColQ anchors AChE at the NMJ remains unknown. We identified five novel mutations at the COOH-terminus of ColQ in seven patients from five families affected with endplate (EP) AChE deficiency. We found that the mutations do not affect the assembly of ColQ with AChE to form asymmetric forms of AChE or impair the interaction of ColQ with perlecan. By contrast, all mutations impair in varied degree the interaction of ColQ to MuSK as well as basement membrane extract (BME) that have no detectable MuSK. Our data confirm that the interaction of ColQ to perlecan and MuSK is crucial for anchoring AChE to the NMJ. In addition, the identified COOH-terminal mutants not only reduce the interaction of ColQ with MuSK, but also diminish the interaction of ColQ with BME. These findings suggest that the impaired attachment of COOH-terminal mutants causing EP AChE deficiency is in part independent of MuSK, and that the COOH-terminus of ColQ may interact with other proteins at the BL. PMID:24281389

  3. Col-OSSOS: Colours of the Outer Solar System Origins Survey

    NASA Astrophysics Data System (ADS)

    Fraser, Wesley Cristopher; Bannister, Michele; Pike, Rosemary; Schwamb, Megan; Marrset, Michael; Kavelaars, JJ; Benecchi, Susan; Delsanti, Audrey; Guilbert-Lepoutre, Audrey; Parker, Alex; Peixinho, Nuno; Vernazza, Peirre; Wang, Shiang-Yu

    2015-11-01

    The surfaces of trans-Neptunian objects (TNOs) are poorly understood. Other than the large objects which exhibit signatures of various ices, very little has been discerned about the compositions of most TNOs. In recent years, some concrete knowledge about the distribution of surface colours of small TNOs has come to light. It is now generally accepted that small TNOs fall into at least three classes of object based on their surface colours and albedo. TNO surface type is also correlated with dynamical class, with certain types of TNO being found primarily in certain regions of the outer Solar System. This correlation presents the intriguing idea that the surfaces of TNOs contain information on more than composition, but as well hold the key to understanding the dynamical processes that lead to the giant planets violently dispersing the protoplanetesimal disk and populating the Kuiper Belt region. It is around this idea that the Col-OSSOS survey is predicated. This 4 year program which started in 2014B is simultaneously using the Gemini-North and Canada-France-Hawaii telescopes to gather near simultaneous u, g, r, and J spectral photometry of all targets in the Outer Solar System Origins Survey (OSSOS) brighter than r’=23.5 (~140 expected). The focus of Col-OSSOS is completeness and consistency, with the same SNR=25 being reached in all bands, for all targets brighter than our depth limit.Col-OSSOS will provide a combined compositional-dynamical map from which key hypotheses about the Solar System's cosmogony can be tested. For example, by mapping the fraction of TNOs with cold-classical like surface colours, we will be able to determine how much of the belt was populated by dynamical scattering versus sweep-up from Neptune. Further, we will be able to constrain the compositional homogeneity of the protoplanetesimal disk. The surfaces of TNOs must reflect that homogeneity; a heterogeneous disk will result in a clumpy colour distribution with many unique types

  4. OsCOL10, a CONSTANS-Like Gene, Functions as a Flowering Time Repressor Downstream of Ghd7 in Rice.

    PubMed

    Tan, Junjie; Jin, Mingna; Wang, Jiachang; Wu, Fuqing; Sheng, Peike; Cheng, Zhijun; Wang, Jiulin; Zheng, Xiaoming; Chen, Liping; Wang, Min; Zhu, Shanshan; Guo, Xiuping; Zhang, Xin; Liu, Xuanming; Wang, Chunming; Wang, Haiyang; Wu, Chuanyin; Wan, Jianmin

    2016-04-01

    Flowering time, or heading date, is a critical agronomic trait that determines the cropping season and regional adaptability, and ultimately grain yield in rice. A number of genes involved in photoperiodic flowering have been cloned and their roles in modulating expression of the flowering genes have been characterized to a certain extent. However, much less is known about the pathway in transmitting the day length response signal(s) to induce transition to reproductive growth. Here, we report a constitutive flowering repressor OsCOL10, which encodes a member of the CONSTANS-like (COL) family. Transgenic rice plants overexpressing OsCOL10 (driven by a strong promoter or by fusing it to the activation domain of VP64) showed delayed flowering time under both short and long days.OsCOL10 is affected by the circadian clock and is preferentially expressed in leaf mesophyll cells; it is localized to the nucleus and has transcriptional activation activity. Further studies show that OsCOL10 represses the expression of theFT-like genes RFT1 and Hd3a through Ehd1. Transcripts of OsCOL10 are more abundant in plants carrying a functional Ghd7 allele or overexpressing Ghd7 than in Ghd7-deficient plants, thus placing OsCOL10 downstream of Ghd7.Taking these findings together, we conclude that OsCOL10 functions as a flowering time repressor that links Ghd7 and Ehd1 in rice. PMID:26872834

  5. Structure of an amidohydrolase, SACOL0085, from methicillin-resistant Staphylococcus aureus COL

    PubMed Central

    Girish, Tavarekere S.; B, Vivek; Colaco, Melwin; Misquith, Sandra; Gopal, B.

    2013-01-01

    Staphylococcus aureus is an opportunistic pathogen that rapidly acquires resistance to frontline antibiotics. The characterization of novel protein targets from this bacterium is thus an important step towards future therapeutic strategies. Here, the crystal structure of an amidohydrolase, SACOL0085, from S. aureus COL is described. SACOL0085 is a member of the M20D family of peptidases. Unlike other M20D peptidases, which are either monomers or dimers, SACOL0085 adopts a butterfly-shaped homotetrameric arrangement with extensive intersubunit interactions. Each subunit of SACOL0085 contains two Mn2+ ions at the active site. A conserved cysteine residue at the active site distinguishes M20D peptidases from other M20 family members. This cysteine, Cys103, serves as bidentate ligand coordinating both Mn2+ ions in SACOL0085. PMID:23385746

  6. Phase-resolved Spectroscopy of the Intermediate Polars -- TV Col and V1223 Sgr

    NASA Astrophysics Data System (ADS)

    Long, K.

    The cataclysmic variables called intermediate polars are characterized by magnetic fields that rip material from an accretion disk and funnel it to a WD that is not phase-locked to the binary period of the system. This is a proposal to use FUSE to conduct a time-resolved spectroscopic study to dissect the emission of two long-period intermediate polars, V1223 Sgr and TV Col, with very different inclination angles. These, along with the short-period high-inclination IP EX Hya (already observed with FUSE), comprise the only IPs with accurate distances derived from HST astrometry. We will isolate emission from the photosphere of the WD, the magnetically dominated accretion curtain, and the accretion stream. Having characterized the emission sources, we will explore the physical conditions in these same regions, and develop an integrated picture of these two intermediate polars.

  7. Investigation on Painting Materials in "Madonna col Bambino e S. Giovannino" by Botticelli

    NASA Astrophysics Data System (ADS)

    Bersani, D.; Lottici, P. P.; Casoli, A.; Ferrari, M.; Lottini, S.; Cauzzi, D.

    A study on the painting materials (pigments and binders) of the famous painting "Madonna col Bambino e S. Giovannino" by Sandro Botticelli, located in the Museo Civico of Piacenza (Italy), was performed before a recent restoration. The painting materials were investigated by the analysis of five millimetric samples taken in damaged regions. The pigments were determined using the micro-Raman spectroscopy, with the 632.8nm line of a He-Ne laser. Despite the strong fluorescence background, the nature of the ground layer (gypsum and anhydrite) and of most pigments (i.e. goethite, lapis lazuli, white lead) was determined. Gas chromatography coupled with mass spectroscopy (GC/MS) was used to determine the organic binder media, and in particular proteinaceous and lipid materials. Egg and animal glue were found, while no siccative oils were detected.

  8. Aberrations of 6q13 Mapped to the COL12A1 Locus in Chondromyxoid Fibroma

    PubMed Central

    Yasuda, Taketoshi; Nishio, Jun; Sumegi, Janos; Kapels, Kayla M.; Althof, Pamela A.; Sawyer, Jeffrey R.; Reith, John D.; Bridge, Julia A.

    2009-01-01

    Chondromyxoid fibroma, a rare benign bone tumor, may be mistaken for chondrosarcoma. Although cytogenetic studies of chondromyxoid fibroma are few, rearrangements of the long arm of chromosome 6 frequently expressed as an inv(6)(p25q13) are prominent. In this study, conventional cytogenetic analysis of 16 chondromyxoid fibroma samples from 14 patients revealed rearrangements of chromosome 6 in ten of eleven clonally abnormal specimens. In addition to 6q13 rearrangements, recurrent 6p25 and 6q25 anomalies were detected. Notably, an identical t(6;9)(q25;q22) translocation was identified in two cases suggesting it represents a distinct translocation of chondromyxoid fibroma. In an effort to further define the aberrant 6q13 breakpoint and identify the molecular consequences, a fluorescence in situ hybridization (FISH)-based positional cloning strategy on chondromyxoid fibroma abnormal metaphase and interphase cells using a series of bacterial and plasmid artificial chromosome (BAC/PAC) probe combinations spanning a 6.1 Mb region was employed. The breakpoint on 6q13 was located within the COL12A1 gene, a collagen gene purportedly involved in another benign bone tumor, subungual exostosis. The findings of this study expand our knowledge of chromosomal alterations in chondromyxoid fibroma, identify COL12A1 as the likely gene candidate within the recurrent 6q13 breakpoint, and provide an alternative approach for detecting 6q13 anomalies in nondividing cells of chondromyxoid fibroma. The latter could potentially be utilized as an adjunct in diagnostically challenging cases. PMID:19648885

  9. Antibiotic resistance due to an unusual ColE1-type replicon plasmid in Aeromonas salmonicida.

    PubMed

    Vincent, Antony T; Emond-Rheault, Jean-Guillaume; Barbeau, Xavier; Attéré, Sabrina A; Frenette, Michel; Lagüe, Patrick; Charette, Steve J

    2016-06-01

    Aeromonas salmonicida subsp. salmonicida is a fish pathogen known to have a rich plasmidome. In the present study, we discovered an isolate of this bacterium bearing an additional unidentified small plasmid. After having sequenced the DNA of that isolate by next-generation sequencing, it appeared that the new small plasmid is a ColE1-type replicon plasmid, named here pAsa7. This plasmid bears a functional chloramphenicol-acetyltransferase-encoding gene (cat-pAsa7) previously unknown in A. salmonicida and responsible for resistance to chloramphenicol. A comparison of pAsa7 with pAsa2, the only known ColE1-type replicon plasmid usually found in A. salmonicida subsp. salmonicida, revealed that even if both plasmids share a high structural similarity, it is still unclear if pAsa7 is a derivative of pAsa2 since they showed several mutations at the nucleotide level. Transcriptomic analysis revealed that the cat-pAsa4 gene, another chloramphenicol-acetyltransferase-encoding gene, found on the large plasmid pAsa4, was significantly more transcribed than cat-pAsa7. This was correlated with a higher chloramphenicol resistance for isolates bearing pAsa4 compared with the one having pAsa7. Finally, a phylogenetic analysis showed that both CAT-pAsa4 and CAT-pAsa7 proteins were in different clusters. The clustering was supported by the identity of residues involved in the catalytic site. In addition, to give a better understanding of the large drug-resistance panel of A. salmonicida, this study reinforces the hypothesis that A. salmonicida subsp. salmonicida is a considerable reservoir for mobile genetic elements such as plasmids. PMID:27028891

  10. A Cratera de Colônia (São Paulo - SP) Aspectos Gerais

    NASA Astrophysics Data System (ADS)

    Varella, Paulo Gomes; Atulim, Regina Auxiliadora

    2006-06-01

    Despite the studies carried out during the last five decades, Colônia crater, situated at the south of São Paulo (SP), is not much known to the Brazilian scientific community (including astronomers) let alone to the population at large. For this paper, we have selected the main characteristics of that crater, such as size, age, geographic location, geological features of the area, and items in favor of its description as an impact crater, since, up to our days, many researchers have been still uncertain as to its origin. We have also established, a comparison between Colônia crater and other similar Brazilian craters, in order to single it out as a very important site for astronomical, paleoclimatic, geological, and geophysical research. It has also been our aim to provide some subsidy to science teachers who wish to approach this subject in the classroom, and stress (emphasize) the importance of this crater as a Brazilian patrimony, considering that this topic, save for a few distinguished exceptions, is not regularly taught at school. Lastly, (finally) we describe, briefly, the current condition of the crater, pointing out the protection initiatives taken by CONDEPHAAT (Conselho de Defesa do Patrimônio Histórico, Artístico, Arqueológico e Turístico do Estado de São Paulo) and the creation of APA (Área de Proteção Ambiental) Capivari-Monos to refrain the disastrous occupation while propitiating a preservative action to protect the important fountainhead area of São Paulo as well as the crater itself.

  11. Frequency of COL4A3/COL4A4 Mutations amongst Families Segregating Glomerular Microscopic Hematuria and Evidence for Activation of the Unfolded Protein Response. Focal and Segmental Glomerulosclerosis Is a Frequent Development during Ageing

    PubMed Central

    Papazachariou, Louiza; Demosthenous, Panayiota; Pieri, Myrtani; Papagregoriou, Gregory; Savva, Isavella; Stavrou, Christoforos; Zavros, Michael; Athanasiou, Yiannis; Ioannou, Kyriakos; Patsias, Charalambos; Panagides, Alexia; Potamitis, Costas; Demetriou, Kyproula; Prikis, Marios; Hadjigavriel, Michael; Kkolou, Maria; Loukaidou, Panayiota; Pastelli, Androulla; Michael, Aristos; Lazarou, Akis; Arsali, Maria; Damianou, Loukas; Goutziamani, Ioanna; Soloukides, Andreas; Yioukas, Lakis; Elia, Avraam; Zouvani, Ioanna; Polycarpou, Polycarpos; Pierides, Alkis; Voskarides, Konstantinos; Deltas, Constantinos

    2014-01-01

    Familial glomerular hematuria(s) comprise a genetically heterogeneous group of conditions which include Alport Syndrome (AS) and thin basement membrane nephropathy (TBMN). Here we investigated 57 Greek-Cypriot families presenting glomerular microscopic hematuria (GMH), with or without proteinuria or chronic kidney function decline, but excluded classical AS. We specifically searched the COL4A3/A4 genes and identified 8 heterozygous mutations in 16 families (28,1%). Eight non-related families featured the founder mutation COL4A3-p.(G1334E). Renal biopsies from 8 patients showed TBMN and focal segmental glomerulosclerosis (FSGS). Ten patients (11.5%) reached end-stage kidney disease (ESKD) at ages ranging from 37-69-yo (mean 50,1-yo). Next generation sequencing of the patients who progressed to ESKD failed to reveal a second mutation in any of the COL4A3/A4/A5 genes, supporting that true heterozygosity for COL4A3/A4 mutations predisposes to CRF/ESKD. Although this could be viewed as a milder and late-onset form of autosomal dominant AS, we had no evidence of ultrastructural features or extrarenal manifestations that would justify this diagnosis. Functional studies in cultured podocytes transfected with wild type or mutant COL4A3 chains showed retention of mutant collagens and differential activation of the unfolded protein response (UPR) cascade. This signifies the potential role of the UPR cascade in modulating the final phenotype in patients with collagen IV nephropathies. PMID:25514610

  12. Frequency of COL4A3/COL4A4 mutations amongst families segregating glomerular microscopic hematuria and evidence for activation of the unfolded protein response. Focal and segmental glomerulosclerosis is a frequent development during ageing.

    PubMed

    Papazachariou, Louiza; Demosthenous, Panayiota; Pieri, Myrtani; Papagregoriou, Gregory; Savva, Isavella; Stavrou, Christoforos; Zavros, Michael; Athanasiou, Yiannis; Ioannou, Kyriakos; Patsias, Charalambos; Panagides, Alexia; Potamitis, Costas; Demetriou, Kyproula; Prikis, Marios; Hadjigavriel, Michael; Kkolou, Maria; Loukaidou, Panayiota; Pastelli, Androulla; Michael, Aristos; Lazarou, Akis; Arsali, Maria; Damianou, Loukas; Goutziamani, Ioanna; Soloukides, Andreas; Yioukas, Lakis; Elia, Avraam; Zouvani, Ioanna; Polycarpou, Polycarpos; Pierides, Alkis; Voskarides, Konstantinos; Deltas, Constantinos

    2014-01-01

    Familial glomerular hematuria(s) comprise a genetically heterogeneous group of conditions which include Alport Syndrome (AS) and thin basement membrane nephropathy (TBMN). Here we investigated 57 Greek-Cypriot families presenting glomerular microscopic hematuria (GMH), with or without proteinuria or chronic kidney function decline, but excluded classical AS. We specifically searched the COL4A3/A4 genes and identified 8 heterozygous mutations in 16 families (28,1%). Eight non-related families featured the founder mutation COL4A3-p.(G1334E). Renal biopsies from 8 patients showed TBMN and focal segmental glomerulosclerosis (FSGS). Ten patients (11.5%) reached end-stage kidney disease (ESKD) at ages ranging from 37-69-yo (mean 50,1-yo). Next generation sequencing of the patients who progressed to ESKD failed to reveal a second mutation in any of the COL4A3/A4/A5 genes, supporting that true heterozygosity for COL4A3/A4 mutations predisposes to CRF/ESKD. Although this could be viewed as a milder and late-onset form of autosomal dominant AS, we had no evidence of ultrastructural features or extrarenal manifestations that would justify this diagnosis. Functional studies in cultured podocytes transfected with wild type or mutant COL4A3 chains showed retention of mutant collagens and differential activation of the unfolded protein response (UPR) cascade. This signifies the potential role of the UPR cascade in modulating the final phenotype in patients with collagen IV nephropathies. PMID:25514610

  13. A sequence that affects the copy number and stability of pSW200 and ColE1.

    PubMed

    Wu, Ying-Chung; Liu, Shih-Tung

    2010-07-01

    Pantoea stewartii SW2 contains 13 plasmids. One of these plasmids, pSW200, has a replicon that resembles that of ColE1. This study demonstrates that pSW200 contains a 9-bp UP element, 5'-AAGATCTTC, which is located immediately upstream of the -35 box in the RNAII promoter. A transcriptional fusion study reveals that substituting this 9-bp sequence reduces the activity of the RNAII promoter by 78%. The same mutation also reduced the number of plasmid copies from 13 to 5, as well as the plasmid stability. When a similar sequence in a ColE1 derivative, pYCW301, is mutated, the copy number of the plasmid also declines from 34 to 16 per cell. Additionally, inserting this 9-bp sequence stabilizes an unstable pSW100 derivative, pSW142K, which also contains a replicon resembling that of ColE1, indicating the importance of this sequence in maintaining the stability of the plasmid. In conclusion, the 9-bp sequence upstream of the -35 box in the RNAII promoter is required for the efficient synthesis of RNAII and maintenance of the stability of the plasmids in the ColE1 family. PMID:20494993

  14. Novel X-linked glomerulopathy associated with a COL4A5 missense mutation in a noncollagenous interruption

    PubMed Central

    Becknell, Brian; Zender, Gloria; Houston, Ronald; Baker, Peter; McBride, Kim L.; Luo, Wentian; Hains, David; Borza, Dorin-Bogdan; Schwaderer, Andrew L.

    2011-01-01

    We report a novel COL4A5 mutation causing rapid progression to end stage renal disease in males despite the absence of clinical and biopsy findings associated with Alport syndrome. Affected males had proteinuria, variable hematuria, early progression to end stage renal disease; and renal biopsy findings which included global and segmental glomerulosclerosis, mesangial hypercellularity and basement membrane immune complex deposition. Exon sequencing of the COL4A5 locus identified a thymine to guanine transversion at nucleotide 665, resulting in a phenylalanine to cysteine missense mutation at codon 222. This mutation was confirmed in 4 affected males and 4 female obligate carriers, but was absent in 6 asymptomatic male family members and 198 unrelated individuals. α5(IV) collagen staining in renal biopsies from affected males was normal. The phenylalanine at position 222 is 100% conserved among vertebrates. This is the first description of a mutation in a non-collagenous interruption associated with severe renal disease, providing evidence for the importance of this structural motif. The range of phenotypes associated with COL4A5 mutations is more diverse than previously realized. COL4A5 mutation analysis should be considered when glomerulonephritis presents in an X-linked inheritance pattern, even with a distinct presentation from Alport syndrome. PMID:20881942

  15. Molecular characterization of CONSTANS-Like (COL) genes in banana (Musa acuminata L. AAA Group, cv. Grand Nain).

    PubMed

    Chaurasia, Akhilesh Kumar; Patil, Hemant Bhagwan; Azeez, Abdul; Subramaniam, Vadakanthara Ramakrishnan; Krishna, Bal; Sane, Aniruddha Prafullachandra; Sane, Prafullachandra Vishnu

    2016-01-01

    The CONSTANS (CO) family is an important regulator of flowering in photoperiod sensitive plants. But information regarding their role in day neutral plants is limited. We report identification of nine Group I type CONSTANS-like (COL) genes of banana and their characterization for their age dependent, diurnal and tissue-specific expression. Our studies show that the Group I genes are conserved in structure to members in other plants. Expression of these genes shows a distinct circadian regulation with a peak during light period. Developmental stage specific expression reveals high level transcript accumulation of two genes, MaCOL3a and MaCOL3b, well before flowering and until the initiation of flowering. A decrease in their transcript levels after initiation of flowering is followed by an increase in transcription of other members that coincides with the continued development of the inflorescence and fruiting. CO binding cis-elements are observed in at least three FT -like genes in banana suggesting possible CO-FT interactions that might regulate flowering. Distinct tissue specific expression patterns are observed for different family members in mature leaves, apical inflorescence, bracts, fruit skin and fruit pulp suggesting possible roles other than flowering. This is the first exhaustive study of the COL genes belonging to Group I of banana. PMID:27186015

  16. Linkage analysis in a family with Stickler syndrome leads to the exclusion of the COL2A1 locus

    SciTech Connect

    Mottes, M.; Zolezzi, F.; Pignatti, P.F.

    1994-09-01

    Hereditary arthro-ophtalmopathy (AO) or Stickler Syndrome (MIM No. 10830) is a dominantly inherited disorder characterized by vitro-retinal degeneration and other connective tissue disturbances. Mutations in the COL2A1 gene, coding for type II collagen chains, have been described in a few patients. The wide spectrum of clinical manifestations is presumably due to genetic heterogeneity, since only about 50% of the Stickler families so far studied show cosegregation of the disease with the COL2A1 locus. We have investigated a large pedigree (19 individuals of whom 9 are affected) in which severe myopia with vitro-retinal degeneration consegregated with joint laxity, recurrent inguinal hernias, and degenerative changes of the hip and the knee. The 3{prime} end COL2A1 VNTR polymorphism was utilized for linkage analysis. In order to get the maximum informativity, we have analyzed the allelic microheterogeneity of this VNTR, due to the repeat sequence variation, by means of a single strand polymorphism. Mendelian inheritance of the different single strands was observed as expected. Discordance of segregation between the disease and the COL2A1 locus was thus established inequivocally in this family.

  17. Mobilization properties of small ColE1-like plasmids carrying kanamycin resistance gene isolated from Salmonella enterica serotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Previously we isolated and characterized various groups of small kanamycin resistance (KanR) ColE1-like plasmids from different serotypes of Salmonella enterica isolates. These plasmids all carried the aph(3)-I gene encoding the aminoglycoside phosphotransferase responsible for the kanam...

  18. Structure of the human type IV collagen COL4A6 gene, which is mutated in Alport syndrome-associated leiomyomatosis

    SciTech Connect

    Zhang, Xu |; Zhou, Jing; Reeders, S.T.

    1996-05-01

    Basement membrane (type IV) collagen, a subfamily of the collagen protein family, is encoded by six distinct genes in mammals. Three of those, COL4A3, COL4A4, and COL4A5, are linked with Alport syndrome (hereditary nephritis). Patients with leimoyomatosis associated with Alport syndrome have been shown to have deletions in the 5{prime} end of the COL4A6 gene, in addition to having deletions in COL4A6. The human COL4A6 gene is reported to be 425 kb as determined by mapping of overlapping YAC clones by probes for its 5{prime} and 3{prime} ends. In the present study we describe the complete exon/intron size pattern of the human COL4A6 gene. The 12 {lambda} phage clones characterized in the study spanned a total of 110 kb, including 85 kb of the actual gene and 25 kb of flanking sequences. The overlapping clones contained all 46 exons of the gene and all introns, except for intron 2. Since the total size of the exons and all introns except for intron 2 is about 85 kb, intron 2 must be about 340 kb. All exons of the gene were assigned to EcoRI restriction fragments to facilitate analysis of the gene in patients with leiomyomatosis associated with Alport syndrome. The exon size pattern of COL4A6 is highly homologous with that of the human and mouse COL4A2 genes, with 27 of the 46 exons of COL4A6 being identical in size between the genes. 42 refs., 2 figs., 3 tabs.

  19. Molecular evolution and phylogenetic analysis of eight COL superfamily genes in group I related to photoperiodic regulation of flowering time in wild and domesticated cotton (Gossypium) species.

    PubMed

    Zhang, Rui; Ding, Jian; Liu, Chunxiao; Cai, Caiping; Zhou, Baoliang; Zhang, Tianzhen; Guo, Wangzhen

    2015-01-01

    Flowering time is an important ecological trait that determines the transition from vegetative to reproductive growth. Flowering time in cotton is controlled by short-day photoperiods, with strict photoperiod sensitivity. As the CO-FT (CONSTANS-FLOWER LOCUS T) module regulates photoperiodic flowering in several plants, we selected eight CONSTANS genes (COL) in group I to detect their expression patterns in long-day and short-day conditions. Further, we individually cloned and sequenced their homologs from 25 different cotton accessions and one outgroup. Finally, we studied their structures, phylogenetic relationship, and molecular evolution in both coding region and three characteristic domains. All the eight COLs in group I show diurnal expression. In the orthologous and homeologous loci, each gene structure in different cotton species is highly conserved, while length variation has occurred due to insertions/deletions in intron and/or exon regions. Six genes, COL2 to COL5, COL7 and COL8, exhibit higher nucleotide diversity in the D-subgenome than in the A-subgenome. The Ks values of 98.37% in all allotetraploid cotton species examined were higher in the A-D and At-Dt comparison than in the A-At and D-Dt comparisons, and the Pearson's correlation coefficient (r) of Ks between A vs. D and At vs. Dt also showed positive, high correlations, with a correlation coefficient of at least 0.797. The nucleotide polymorphism in wild species is significantly higher compared to G. hirsutum and G. barbadense, indicating a genetic bottleneck associated with the domesticated cotton species. Three characteristic domains in eight COLs exhibit different evolutionary rates, with the CCT domain highly conserved, while the B-box and Var domain much more variable in allotetraploid species. Taken together, COL1, COL2 and COL8 endured greater selective pressures during the domestication process. The study improves our understanding of the domestication-related genes/traits during cotton

  20. Knockdown of col22a1 gene in zebrafish induces a muscular dystrophy by disruption of the myotendinous junction.

    PubMed

    Charvet, Benjamin; Guiraud, Alexandre; Malbouyres, Marilyne; Zwolanek, Daniela; Guillon, Emilie; Bretaud, Sandrine; Monnot, Catherine; Schulze, Jörg; Bader, Hannah L; Allard, Bruno; Koch, Manuel; Ruggiero, Florence

    2013-11-01

    The myotendinous junction (MTJ) is the major site of force transfer in skeletal muscle, and defects in its structure correlate with a subset of muscular dystrophies. Col22a1 encodes the MTJ component collagen XXII, the function of which remains unknown. Here, we have cloned and characterized the zebrafish col22a1 gene and conducted morpholino-based loss-of-function studies in developing embryos. We showed that col22a1 transcripts localize at muscle ends when the MTJ forms and that COLXXII protein integrates the junctional extracellular matrix. Knockdown of COLXXII expression resulted in muscular dystrophy-like phenotype, including swimming impairment, curvature of embryo trunk/tail, strong reduction of twitch-contraction amplitude and contraction-induced muscle fiber detachment, and provoked significant activation of the survival factor Akt. Electron microscopy and immunofluorescence studies revealed that absence of COLXXII caused a strong reduction of MTJ folds and defects in myoseptal structure. These defects resulted in reduced contractile force and susceptibility of junctional extracellular matrix to rupture when subjected to repeated mechanical stress. Co-injection of sub-phenotypic doses of morpholinos against col22a1 and genes of the major muscle linkage systems showed a synergistic gene interaction between col22a1 and itga7 (α7β1 integrin) that was not observed with dag1 (dystroglycan). Finally, pertinent to a conserved role in humans, the dystrophic phenotype was rescued by microinjection of recombinant human COLXXII. Our findings indicate that COLXXII contributes to the stabilization of myotendinous junctions and strengthens skeletal muscle attachments during contractile activity. PMID:24131632

  1. PTH stimulated growth and decreased Col-X deposition are phosphotidylinositol-3,4,5 triphosphate kinase and mitogen activating protein kinase dependent in avian sterna.

    PubMed

    Harrington, Erik Kern; Coon, David J; Kern, Matthew F; Svoboda, Kathy K H

    2010-02-01

    Type X collagen (Col-X) deposition is a marker of terminal differentiation during chondrogenesis, in addition to appositional growth and apoptosis. The parathyroid hormone/parathyroid hormone related peptide (PTH/PTHrP) receptor, or PPR, is a G-Protein coupled receptor (GPCR), which activates several downstream pathways, moderating chondrocyte differentiation, including suppression of Col-X deposition. An Avian sterna model was used to analyze the PPR GPCR downstream kinase role in growth rate and extracellular matrix (ECM) including Col-II, IX, and X. Phosphatidylinositol kinase (PI3K), mitogen activating protein kinase (MAPK) and protein kinase A (PKA) were inhibited with specific established inhibitors LY294002, PD98059, and H89, respectively to test the hypothesis that they could reverse/inhibit the PTH/PTHrP pathway. Excised E14 chick sterna were PTH treated with or without an inhibitor and compared to controls. Sternal length was measured every 24 hr. Cultured sterna were immuno-stained using specific antibodies for Col-II, IX, or X and examined via confocal microscopy. Increased growth in PTH-treated sterna was MAPK, PI3K, and PKA dose dependent, suggesting growth was regulated through multiple pathways. Col-X deposition was rescued in PTH-treated sterna in the presence of PI3K or MAPK inhibitors, but not with the PKA inhibitor. All three inhibitors moderately disrupted Col-II and Col-IX deposition. These results suggest that PTH can activate multiple pathways during chondrocyte differentiation. PMID:19957341

  2. Linkage mapping of the gene for Type III collagen (COL3A1) to human chromosome 2q using a VNTR polymorphism

    SciTech Connect

    Tiller, G.E.; Polumbo, P.A.; Summar, M.L. )

    1994-03-15

    The gene for the [alpha]1(III) chain of type III collagen, COL3A1, has been previously mapped to human chromosome 2q24.3-q31 by in situ hybridization. Physical mapping by pulsed-field gel electrophoresis has demonstrated that COL3A1 lies within 35 kb of COL5A2. The authors genotyped the CEPH families at the COL3A2 locus using a pentanucleotide repeat polymorphism within intron 25. They demonstrated significant linkage to 18 anonymous markers as well as the gene for carbamyl phosphate synthetase (CPSI), which had been previously mapped to this region. No recombination was seen between COL3A1 and COL5A2 (Z = 9.93 at [theta] = 0) or D2S24 (Z = 10.55 at [theta] = 0). The locus order is (D2S32-D2S138-D2S148)-(D2S24-COL5A2-COL3A1)-(D2S118-D2S161), with odds of 1:2300 for the next most likely order. These relationships are consistent with the physical mapping of COL3A1 to the distal portion of 2q and place it proximal to CPSI by means of multipoint analysis. These linkage relationships should prove useful in further studies of Ehlers-Danlos syndrome type IV and carbamyl phosphate synthetase I deficiency and provide an additional framework for localizing other genes in this region. 13 refs., 2 figs., 1 tab.

  3. Neuromuscular junction immaturity and muscle atrophy are hallmarks of the ColQ-deficient mouse, a model of congenital myasthenic syndrome with acetylcholinesterase deficiency.

    PubMed

    Sigoillot, Séverine M; Bourgeois, Francine; Karmouch, Jennifer; Molgó, Jordi; Dobbertin, Alexandre; Chevalier, Catherine; Houlgatte, Rémi; Léger, Jean; Legay, Claire

    2016-06-01

    The collagen ColQ anchors acetylcholinesterase (AChE) in the synaptic cleft of the neuromuscular junction (NMJ). It also binds MuSK and perlecan/dystroglycan, 2 signaling platforms of the postsynaptic domain. Mutations in ColQ cause a congenital myasthenic syndrome (CMS) with AChE deficiency. Because the absence of AChE does not fully explain the complexity of the syndrome and there is no curative treatment for the disease, we explored additional potential targets of ColQ by conducting a large genetic screening of ColQ-deficient mice, a model for CMS with AChE deficiency, and analyzed their NMJ and muscle phenotypes. We demonstrated that ColQ controls the development and the maturation of the postsynaptic domain by regulating synaptic gene expression. Notably, ColQ deficiency leads to an up-regulation of the 5 subunits of the nicotinic acetylcholine receptor (AChR), leading to mixed mature and immature AChRs at the NMJ of adult mice. ColQ also regulates the expression of extracellular matrix (ECM) components. However, whereas the ECM mRNAs were down-regulated in vitro, compensation seemed to occur in vivo to maintain normal levels of these mRNAs. Finally, ColQ deficiency leads to a general atrophic phenotype and hypoplasia that affect fast muscles. This study points to new specific hallmarks for this CMS.-Sigoillot, S. M., Bourgeois, F., Karmouch, J., Molgó, J., Dobbertin, A., Chevalier, C., Houlgatte, R., Léger, J., Legay, C. Neuromuscular junction immaturity and muscle atrophy are hallmarks of the ColQ-deficient mouse, a model of congenital myasthenic syndrome with acetylcholinesterase deficiency. PMID:26993635

  4. The procollagen type III, alpha 1 (COL3A1) gene first intron expresses poly-A+ RNA corresponding to multiple ESTs and putative miRNAs.

    PubMed

    Sterling, Kenneth M

    2011-02-01

    The mouse COL3A1 first intron is 9684 bp. RNA's of approximately 1.6 and 3.0 kb were detected by Northern hybridization analysis of poly-A RNA from fetal mice and total RNA from suckling and adult mouse intestine using (32)P-labeled, anti-sense RNA synthesized from a mouse COL3A1 first intron, 5 prime region, 5.4 kb Xba I fragment (1655-7030 bp), recombinant plasmid (pPI5.4x). Expression of the 1.6 and 3.0 kb RNA's was significantly reduced in adult mouse intestine, indicating that these RNAs are developmentally regulated. "BLAST" analysis indicated that the mouse first intron 5 prime sequence has 94-100% identity to 13 mouse ESTs. These mouse first intron EST's lie within the 5.4 Xba I fragment of the mouse COL3A1 first intron. Two of the mouse first intron EST's have significant identity to known miRNA, mature sequences, mmu-miR-466f-3P, mmu-miR-1187, and mmu-miR-574-5P as well as others. Predicted targets for mmu-miR-466f-3P include COL1A1, COL19A1, COL11A2, COL4A1, and COL4A5 indicating that COL3A1 intronic miRNAs may regulate the expression of other collagen genes in development. PMID:21268075

  5. Novel form of X-linked nonsyndromic hearing loss with cochlear malformation caused by a mutation in the type IV collagen gene COL4A6

    PubMed Central

    Rost, Simone; Bach, Elisa; Neuner, Cordula; Nanda, Indrajit; Dysek, Sandra; Bittner, Reginald E; Keller, Alexander; Bartsch, Oliver; Mlynski, Robert; Haaf, Thomas; Müller, Clemens R; Kunstmann, Erdmute

    2014-01-01

    Hereditary hearing loss is the most common human sensorineural disorder. Genetic causes are highly heterogeneous, with mutations detected in >40 genes associated with nonsyndromic hearing loss, to date. Whereas autosomal recessive and autosomal dominant inheritance is prevalent, X-linked forms of nonsyndromic hearing impairment are extremely rare. Here, we present a Hungarian three-generation family with X-linked nonsyndromic congenital hearing loss and the underlying genetic defect. Next-generation sequencing and subsequent segregation analysis detected a missense mutation (c.1771G>A, p.Gly591Ser) in the type IV collagen gene COL4A6 in all affected family members. Bioinformatic analysis and expression studies support this substitution as being causative. COL4A6 encodes the alpha-6 chain of type IV collagen of basal membranes, which forms a heterotrimer with two alpha-5 chains encoded by COL4A5. Whereas mutations in COL4A5 and contiguous X-chromosomal deletions involving COL4A5 and COL4A6 are associated with X-linked Alport syndrome, a nephropathy associated with deafness and cataract, mutations in COL4A6 alone have not been related to any hereditary disease so far. Moreover, our index patient and other affected family members show normal renal and ocular function, which is not consistent with Alport syndrome, but with a nonsyndromic type of hearing loss. In situ hybridization and immunostaining demonstrated expression of the COL4A6 homologs in the otic vesicle of the zebrafish and in the murine inner ear, supporting its role in normal ear development and function. In conclusion, our results suggest COL4A6 as being the fourth gene associated with X-linked nonsyndromic hearing loss. PMID:23714752

  6. Novel form of X-linked nonsyndromic hearing loss with cochlear malformation caused by a mutation in the type IV collagen gene COL4A6.

    PubMed

    Rost, Simone; Bach, Elisa; Neuner, Cordula; Nanda, Indrajit; Dysek, Sandra; Bittner, Reginald E; Keller, Alexander; Bartsch, Oliver; Mlynski, Robert; Haaf, Thomas; Müller, Clemens R; Kunstmann, Erdmute

    2014-02-01

    Hereditary hearing loss is the most common human sensorineural disorder. Genetic causes are highly heterogeneous, with mutations detected in >40 genes associated with nonsyndromic hearing loss, to date. Whereas autosomal recessive and autosomal dominant inheritance is prevalent, X-linked forms of nonsyndromic hearing impairment are extremely rare. Here, we present a Hungarian three-generation family with X-linked nonsyndromic congenital hearing loss and the underlying genetic defect. Next-generation sequencing and subsequent segregation analysis detected a missense mutation (c.1771G>A, p.Gly591Ser) in the type IV collagen gene COL4A6 in all affected family members. Bioinformatic analysis and expression studies support this substitution as being causative. COL4A6 encodes the alpha-6 chain of type IV collagen of basal membranes, which forms a heterotrimer with two alpha-5 chains encoded by COL4A5. Whereas mutations in COL4A5 and contiguous X-chromosomal deletions involving COL4A5 and COL4A6 are associated with X-linked Alport syndrome, a nephropathy associated with deafness and cataract, mutations in COL4A6 alone have not been related to any hereditary disease so far. Moreover, our index patient and other affected family members show normal renal and ocular function, which is not consistent with Alport syndrome, but with a nonsyndromic type of hearing loss. In situ hybridization and immunostaining demonstrated expression of the COL4A6 homologs in the otic vesicle of the zebrafish and in the murine inner ear, supporting its role in normal ear development and function. In conclusion, our results suggest COL4A6 as being the fourth gene associated with X-linked nonsyndromic hearing loss. PMID:23714752

  7. ColE1-Plasmid Production in Escherichia coli: Mathematical Simulation and Experimental Validation

    PubMed Central

    Freudenau, Inga; Lutter, Petra; Baier, Ruth; Schleef, Martin; Bednarz, Hanna; Lara, Alvaro R.; Niehaus, Karsten

    2015-01-01

    Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the past years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3) and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ) by using ordinary differential equations and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII) as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN), and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI- and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ± 0.7 and 34 ± 7 RNAI molecules per cell and 0.44 ± 0.1 and 3 ± 0.9 RNAII molecules per cell. The determined PCNs averaged between 46 ± 26 and 48 ± 30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ± 203 and 1086 ± 298 RNAI molecules per cell and 22 ± 2 and 75 ± 10 RNAII molecules per cell with an averaged PCN of 1514 ± 1301 and 5806 ± 4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the plasmid DNA production. PMID:26389114

  8. Trabecular Bone Deterioration in col9a1 +/− Mice Associated With Enlarged Osteoclasts Adhered to Collagen IX–Deficient Bone

    PubMed Central

    Wang, Chiachien Jake; Iida, Keisuke; Egusa, Hiroshi; Hokugo, Akishige; Jewett, Anahid; Nishimura, Ichiro

    2008-01-01

    Introduction Short collagen IX, the exclusive isoform expressed by osteoblasts, is synthesized through alternative transcription of the col9a1 gene. The function of short collagen IX in bone was characterized in col9a1-null mutant mice. Materials and Methods Trabecular bone morphometry of lumbar bones and tibias was evaluated by μCT and nondecalcified histology. Osteoblastic and osteoclastic activities were evaluated by PCR- and microarray-based gene expression assays and TRACP-5b and C-terminal telopeptide (CTX) assays, as well as in vitro using bone marrow stromal cells and splenocytes. The effect of col9a1+/− mutation on osteoclast morphology was evaluated using RAW264.7-derived osteoclastic cells cultured on the mutant or wildtype calvarial bone substrates. Results Col9a1 knockout mutation caused little effects on the skeletal development; however, young adult female col9a1 −/− and col9a1 +/− mice exhibited significant loss of trabecular bone. The trabecular bone architecture was progressively deteriorated in both male and female heterozygous col9a1 +/− mice while aging. The aged mutant mice also exhibited signs of thoracic kyphosis and weight loss, resembling the clinical signs of osteoporosis. The col9a1 +/− osteoblasts synthesized short col9a1 transcripts at decreased rates. Whereas bone formation activities in vitro and in vivo were not affected, the mutant osteoblast expressed the elevated ratio of RANKL/osteoprotegerin. Increased serum TRACP-5b and CTX levels were found in col9a1 +/− mice, whose bone surface was associated with osteoclastic cells that were abnormally flattened and enlarged. The mutant and wildtype splenocytes underwent similar osteoclastogenesis in vitro; however, RAW264.7-derived osteoclastic cells, when cultured on the col9a1 +/− calvaria, widely spread over the bone surface and formed large resorption pits. The surface of col9a1 +/− calvaria was found to lack the typical nanotopography. Conclusions The mineralized

  9. A repeat sequence causes competition of ColE1-type plasmids.

    PubMed

    Lin, Mei-Hui; Fu, Jen-Fen; Liu, Shih-Tung

    2013-01-01

    Plasmid pSW200 from Pantoea stewartii contains 41 copies of 15-bp repeats and has a replicon that is homologous to that of ColE1. Although deleting the repeats (pSW207) does not change the copy number and stability of the plasmid. The plasmid becomes unstable and is rapidly lost from the host when a homoplasmid with the repeats (pSW201) is present. Deleting the repeats is found to reduce the transcriptional activity of RNAIp and RNAIIp by about 30%, indicating that the repeats promote the transcription of RNAI and RNAII, and how the RNAI that is synthesized by pSW201 inhibits the replication of pSW207. The immunoblot analysis herein demonstrates that RNA polymerase β subunit and σ(70) in the lysate from Escherichia coli MG1655 bind to a biotin-labeled DNA probe that contains the entire sequence of the repeat region. Electrophoretic mobility shift assay also reveals that purified RNA polymerase shifts a DNA probe that contains four copies of the repeats. These results thus obtained reveal that RNA polymerase holoenzyme binds to the repeats. The repeats also exchange RNA polymerase with RNAIp and RNAIIp in vitro, revealing the mechanism by which the transcription is promoted. This investigation elucidates a mechanism by which a plasmid prevents the invasion of an incompatible plasmid and maintains its stability in the host cell during evolution. PMID:23613898

  10. AUDIOLOGICAL FINDINGS IN OTOSPONDYLOMEGAEPIPHYSEAL DYSPLASIA (OSMED) ASSOCIATED WITH A NOVEL MUTATION IN COL11A2

    PubMed Central

    Tokgöz-Yilmaz, Suna; Şahli, Sanem; Fitoz, Suat; Sennaroğlu, Gonca; Tekin, Mustafa

    2011-01-01

    Objective The aim of the study was to assess the audiological findings of a 4-year-old child with a homozygous COL11A2 mutation and to point out the role of continuous follow-ups in children with craniofacial syndromes after the newborn hearing screening. Methods A 4-year-old boy with otospondylomegaepiphyseal dysplasia (OSMED) was followed up after birth for hearing loss. Transient Otoacoustic Emissions (TEOAE’s), Distortion Product Otoacoustic Emissions (DPOAE’s), Automated and Clinical Auditory Brainstem Response (AABR and ABR) measurements, Visual Reinforcement Audiometry, immitansmetric measurements and hearing threshold measurements were performed for audiological evaluation. Results The patient developed sensorineural hearing loss at 11 months of age while his hearing was normal at birth. Because of auditory-verbal training with hearing aids started at 20 months of age, he now has normal verbal communication with his peers. Conclusions This study clearly demonstrates that hearing loss developes in infancy in patients with OSMED and underscrores the importance of contunied hearing screening beyond newborn period for early intervention of hearing impairment and communication problems. PMID:21208667

  11. Unique mutation portraits and frequent COL2A1 gene alteration in chondrosarcoma

    PubMed Central

    Totoki, Yasushi; Yoshida, Akihiko; Hosoda, Fumie; Nakamura, Hiromi; Hama, Natsuko; Ogura, Koichi; Yoshida, Aki; Fujiwara, Tomohiro; Arai, Yasuhito; Toguchida, Junya; Tsuda, Hitoshi; Miyano, Satoru; Kawai, Akira

    2014-01-01

    Chondrosarcoma is the second most frequent malignant bone tumor. However, the etiological background of chondrosarcomagenesis remains largely unknown, along with details on molecular alterations and potential therapeutic targets. Massively parallel paired-end sequencing of whole genomes of 10 primary chondrosarcomas revealed that the process of accumulation of somatic mutations is homogeneous irrespective of the pathological subtype or the presence of IDH1 mutations, is unique among a range of cancer types, and shares significant commonalities with that of prostate cancer. Clusters of structural alterations localized within a single chromosome were observed in four cases. Combined with targeted resequencing of additional cartilaginous tumor cohorts, we identified somatic alterations of the COL2A1 gene, which encodes an essential extracellular matrix protein in chondroskeletal development, in 19.3% of chondrosarcoma and 31.7% of enchondroma cases. Epigenetic regulators (IDH1 and YEATS2) and an activin/BMP signal component (ACVR2A) were recurrently altered. Furthermore, a novel FN1-ACVR2A fusion transcript was observed in both chondrosarcoma and osteochondromatosis cases. With the characteristic accumulative process of somatic changes as a background, molecular defects in chondrogenesis and aberrant epigenetic control are primarily causative of both benign and malignant cartilaginous tumors. PMID:25024164

  12. Genetic and structural analysis of the ColE1 Rop (Rom) protein.

    PubMed Central

    Castagnoli, L; Scarpa, M; Kokkinidis, M; Banner, D W; Tsernoglou, D; Cesareni, G

    1989-01-01

    Repressor of primer (Rop) is a small dimeric protein that participates in the mechanism that controls the copy number of plasmid of the ColE1 family by increasing the affinity between two complementary RNAs. The Rop dimer is a bundle of four tightly packed alpha-helices that are held together by hydrophobic interactions. We have systematically altered, by site directed mutagenesis, most of the solvent exposed amino acids of the Rop bundle and we have identified the alterations that cause a decrease of the activity of the regulatory molecule. We conclude that Rop folding is rather insensitive to amino acid substitutions and to other mutations as drastic as deletions and insertions. Looking along the 2-fold symmetry axis the amino acid side chains whose alterations affect the function of Rop are all located on one side of the molecule. Furthermore they are clustered at the extremities of the alpha-helix bundle, the only exception being the aromatic ring of Phe-14. Images PMID:2721494

  13. Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB

    PubMed Central

    Georgiadis, Christos; Syed, Farhatullah; Petrova, Anastasia; Abdul-Wahab, Alya; Lwin, Su M.; Farzaneh, Farzin; Chan, Lucas; Ghani, Sumera; Fleck, Roland A.; Glover, Leanne; McMillan, James R.; Chen, Mei; Thrasher, Adrian J.; McGrath, John A.; Di, Wei-Li; Qasim, Waseem

    2016-01-01

    Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen, a protein essential for anchoring fibril formation at the dermal-epidermal junction. Whereas allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection and support sustained delivery of type VII collagen at the dermal-epidermal junction. We demonstrate the feasibility of such an approach using a therapeutic grade, self-inactivating-lentiviral vector, encoding codon-optimized COL7A1, to transduce RDEB fibroblasts under conditions suitable for clinical application. Expression and secretion of type VII collagen was confirmed with transduced cells exhibiting supranormal levels of protein expression, and ex vivo migration of fibroblasts was restored in functional assays. Gene-modified RDEB fibroblasts also deposited type VII collagen at the dermal-epidermal junction of human RDEB skin xenografts placed on NOD-scid IL2Rgammanull recipients, with reconstruction of human epidermal structure and regeneration of anchoring fibrils at the dermal-epidermal junction. Fibroblast-mediated restoration of protein and structural defects in this RDEB model strongly supports proposed therapeutic applications in man. PMID:26763448

  14. Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB.

    PubMed

    Georgiadis, Christos; Syed, Farhatullah; Petrova, Anastasia; Abdul-Wahab, Alya; Lwin, Su M; Farzaneh, Farzin; Chan, Lucas; Ghani, Sumera; Fleck, Roland A; Glover, Leanne; McMillan, James R; Chen, Mei; Thrasher, Adrian J; McGrath, John A; Di, Wei-Li; Qasim, Waseem

    2016-01-01

    Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen, a protein essential for anchoring fibril formation at the dermal-epidermal junction. Whereas allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection and support sustained delivery of type VII collagen at the dermal-epidermal junction. We demonstrate the feasibility of such an approach using a therapeutic grade, self-inactivating-lentiviral vector, encoding codon-optimized COL7A1, to transduce RDEB fibroblasts under conditions suitable for clinical application. Expression and secretion of type VII collagen was confirmed with transduced cells exhibiting supranormal levels of protein expression, and ex vivo migration of fibroblasts was restored in functional assays. Gene-modified RDEB fibroblasts also deposited type VII collagen at the dermal-epidermal junction of human RDEB skin xenografts placed on NOD-scid IL2Rgamma(null) recipients, with reconstruction of human epidermal structure and regeneration of anchoring fibrils at the dermal-epidermal junction. Fibroblast-mediated restoration of protein and structural defects in this RDEB model strongly supports proposed therapeutic applications in man. PMID:26763448

  15. Improved genetic counseling in Alport syndrome by new variants of COL4A5 gene.

    PubMed

    Fernandez-Rosado, Francisco; Campos, Ana; Alvarez-Cubero, Maria Jesus; Ruiz, Ana; Entrala-Bernal, Carmen

    2015-07-01

    There are current requirements of using genetic databases for offering a better genetic assistance to patients of some syndromes, especially those with X-linked heredity patterns (like Alport Syndrome) for the high probability of having descendants affected by the disease. We describe the first reported case of COL4A5 gene missense c.1499 G>T mutation in a 16-year-old girl confirmed to be affected by Alport Syndrome after genetic counseling. Next Generation Sequencing procedures let discover this mutation and offer an accurate clinical treatment to this patient. Current scientific understanding of genetic syndromes suggests the high importance of updated databases and the inclusion of Variant of Unknown Significance related to clinical cases. All of this updating could enable patients to have a better opportunity of diagnosis and having genetic and clinical counseling. This event is even more important in women planning to start a family to have correct genetic counseling regarding the risk posed to offspring, and allowing the decision to undergo prenatal testing. PMID:26063487

  16. Identification of a Novel Mutation in the COL2A1 Gene in a Chinese Family with Spondyloepiphyseal Dysplasia Congenita

    PubMed Central

    Huang, Xiangjun; Deng, Xiong; Xu, Hongbo; Wu, Song; Yuan, Lamei; Yang, Zhijian; Yang, Yan; Deng, Hao

    2015-01-01

    Spondyloepiphyseal dysplasia congenita (SEDC) is an autosomal dominant chondrodysplasia characterized by disproportionate short-trunk dwarfism, skeletal and vertebral deformities. Exome sequencing and Sanger sequencing were performed in a Chinese Han family with typical SEDC, and a novel mutation, c.620G>A (p.Gly207Glu), in the collagen type II alpha-1 gene (COL2A1) was identified. The mutation may impair protein stability, and lead to dysfunction of type II collagen. Family-based study suggested that the mutation is a de novo mutation. Our study extends the mutation spectrum of SEDC and confirms genotype-phenotype relationship between mutations at glycine in the triple helix of the alpha-1(II) chains of the COL2A1 and clinical findings of SEDC, which may be helpful in the genetic counseling of patients with SEDC. PMID:26030151

  17. Novel mutations confirm that COL11A2 is responsible for autosomal recessive non-syndromic hearing loss DFNB53.

    PubMed

    Chakchouk, Imen; Grati, M'hamed; Bademci, Guney; Bensaid, Mariem; Ma, Qi; Chakroun, Amine; Foster, Joseph; Yan, Denise; Duman, Duygu; Diaz-Horta, Oscar; Ghorbel, Abdelmonem; Mittal, Rahul; Farooq, Amjad; Tekin, Mustafa; Masmoudi, Saber; Liu, Xue Zhong

    2015-08-01

    Hearing loss (HL) is a major public health issue. It is clinically and genetically heterogeneous.The identification of the causal mutation is important for early diagnosis, clinical follow-up, and genetic counseling. HL due to mutations in COL11A2, encoding collagen type XI alpha-2, can be non-syndromic autosomal-dominant or autosomal-recessive, and also syndromic as in Otospondylomegaepiphyseal Dysplasia, Stickler syndrome type III, and Weissenbacher-Zweymuller syndrome. However, thus far only one mutation co-segregating with autosomal recessive non-syndromic hearing loss (ARNSHL) in a single family has been reported. In this study, whole exome sequencing of two consanguineous families with ARNSHL from Tunisia and Turkey revealed two novel causative COL11A2 mutations, c.109G > T (p.Ala37Ser) and c.2662C > A (p.Pro888Thr). The variants identified co-segregated with deafness in both families. All homozygous individuals in those families had early onset profound hearing loss across all frequencies without syndromic findings. The variants are predicted to be damaging the protein function. The p.Pro888Thr mutation affects a -Gly-X-Y- triplet repeat motif. The novel p.Ala37Ser is the first missense mutation located in the NC4 domain of the COL11A2 protein. Structural model suggests that this mutation will likely obliterate, or at least partially compromise, the ability of NC4 domain to interact with its cognate ligands. In conclusion, we confirm that COL11A2 mutations cause ARNSHL and broaden the mutation spectrum that may shed new light on genotype-phenotype correlation for the associated phenotypes and clinical follow-up. PMID:25633957

  18. Novel mutations confirm that COL11A2 is responsible for autosomal recessive non-syndromic hearing loss DFNB53

    PubMed Central

    Chakchouk, Imen; Grati, M’hamed; Bademci, Guney; Bensaid, Mariem; Ma, Qi; Chakroun, Amine; Foster, Joseph; Yan, Denise; Duman, Duygu; Diaz-Horta, Oscar; Ghorbel, Abdelmonem; Mittal, Rahul; Farooq, Amjad; Tekin, Mustafa

    2015-01-01

    Hearing loss (HL) is a major public health issue. It is clinically and genetically heterogeneous. The identification of the causal mutation is important for early diagnosis, clinical follow-up, and genetic counseling. HL due to mutations in COL11A2, encoding collagen type XI alpha-2, can be non-syndromic autosomal-dominant or autosomal-recessive, and also syndromic as in Otospondylomegaepiphyseal Dysplasia, Stickler syndrome type III, and Weissenbacher–Zweymuller syndrome. However, thus far only one mutation co-segregating with autosomal recessive non-syndromic hearing loss (ARNSHL) in a single family has been reported. In this study, whole exome sequencing of two consanguineous families with ARNSHL from Tunisia and Turkey revealed two novel causative COL11A2 mutations, c.109G > T (p.Ala37Ser) and c.2662C > A (p.Pro888Thr). The variants identified co-segregated with deafness in both families. All homozygous individuals in those families had early onset profound hearing loss across all frequencies without syndromic findings. The variants are predicted to be damaging the protein function. The p.Pro888Thr mutation affects a -Gly-X–Y- triplet repeat motif. The novel p.Ala37Ser is the first missense mutation located in the NC4 domain of the COL11A2 protein. Structural model suggests that this mutation will likely obliterate, or at least partially compromise, the ability of NC4 domain to interact with its cognate ligands. In conclusion, we confirm that COL11A2 mutations cause ARNSHL and broaden the mutation spectrum that may shed new light on genotype–phenotype correlation for the associated phenotypes and clinical follow-up. PMID:25633957

  19. Recessive Mutations in the α3 (VI) Collagen Gene COL6A3 Cause Early-Onset Isolated Dystonia

    PubMed Central

    Zech, Michael; Lam, Daniel D.; Francescatto, Ludmila; Schormair, Barbara; Salminen, Aaro V.; Jochim, Angela; Wieland, Thomas; Lichtner, Peter; Peters, Annette; Gieger, Christian; Lochmüller, Hanns; Strom, Tim M.; Haslinger, Bernhard; Katsanis, Nicholas; Winkelmann, Juliane

    2015-01-01

    Isolated dystonia is a disorder characterized by involuntary twisting postures arising from sustained muscle contractions. Although autosomal-dominant mutations in TOR1A, THAP1, and GNAL have been found in some cases, the molecular mechanisms underlying isolated dystonia are largely unknown. In addition, although emphasis has been placed on dominant isolated dystonia, the disorder is also transmitted as a recessive trait, for which no mutations have been defined. Using whole-exome sequencing in a recessive isolated dystonia-affected kindred, we identified disease-segregating compound heterozygous mutations in COL6A3, a collagen VI gene associated previously with muscular dystrophy. Genetic screening of a further 367 isolated dystonia subjects revealed two additional recessive pedigrees harboring compound heterozygous mutations in COL6A3. Strikingly, all affected individuals had at least one pathogenic allele in exon 41, including an exon-skipping mutation that induced an in-frame deletion. We tested the hypothesis that disruption of this exon is pathognomonic for isolated dystonia by inducing a series of in-frame deletions in zebrafish embryos. Consistent with our human genetics data, suppression of the exon 41 ortholog caused deficits in axonal outgrowth, whereas suppression of other exons phenocopied collagen deposition mutants. All recessive mutation carriers demonstrated early-onset segmental isolated dystonia without muscular disease. Finally, we show that Col6a3 is expressed in neurons, with relevant mRNA levels detectable throughout the adult mouse brain. Taken together, our data indicate that loss-of-function mutations affecting a specific region of COL6A3 cause recessive isolated dystonia with underlying neurodevelopmental deficits and highlight the brain extracellular matrix as a contributor to dystonia pathogenesis. PMID:26004199

  20. Recessive mutations in the α3 (VI) collagen gene COL6A3 cause early-onset isolated dystonia.

    PubMed

    Zech, Michael; Lam, Daniel D; Francescatto, Ludmila; Schormair, Barbara; Salminen, Aaro V; Jochim, Angela; Wieland, Thomas; Lichtner, Peter; Peters, Annette; Gieger, Christian; Lochmüller, Hanns; Strom, Tim M; Haslinger, Bernhard; Katsanis, Nicholas; Winkelmann, Juliane

    2015-06-01

    Isolated dystonia is a disorder characterized by involuntary twisting postures arising from sustained muscle contractions. Although autosomal-dominant mutations in TOR1A, THAP1, and GNAL have been found in some cases, the molecular mechanisms underlying isolated dystonia are largely unknown. In addition, although emphasis has been placed on dominant isolated dystonia, the disorder is also transmitted as a recessive trait, for which no mutations have been defined. Using whole-exome sequencing in a recessive isolated dystonia-affected kindred, we identified disease-segregating compound heterozygous mutations in COL6A3, a collagen VI gene associated previously with muscular dystrophy. Genetic screening of a further 367 isolated dystonia subjects revealed two additional recessive pedigrees harboring compound heterozygous mutations in COL6A3. Strikingly, all affected individuals had at least one pathogenic allele in exon 41, including an exon-skipping mutation that induced an in-frame deletion. We tested the hypothesis that disruption of this exon is pathognomonic for isolated dystonia by inducing a series of in-frame deletions in zebrafish embryos. Consistent with our human genetics data, suppression of the exon 41 ortholog caused deficits in axonal outgrowth, whereas suppression of other exons phenocopied collagen deposition mutants. All recessive mutation carriers demonstrated early-onset segmental isolated dystonia without muscular disease. Finally, we show that Col6a3 is expressed in neurons, with relevant mRNA levels detectable throughout the adult mouse brain. Taken together, our data indicate that loss-of-function mutations affecting a specific region of COL6A3 cause recessive isolated dystonia with underlying neurodevelopmental deficits and highlight the brain extracellular matrix as a contributor to dystonia pathogenesis. PMID:26004199

  1. COL4A4 gene study of a European population: description of new mutations causing autosomal dominant Alport syndrome

    PubMed Central

    Rosado, Consolación; Bueno, Elena; Felipe, Carmen; González-Sarmiento, Rogelio

    2014-01-01

    Background: Autosomal forms of Alport syndrome represent 20% of all patients (15% recessive and 5% dominant). They are caused by mutations in the COL4A3 and COL4A4 genes, which encode a-3 and a-4 collagen IV chains of the glomerular basement membrane, cochlea and eye. Thin basement membrane nephropathy may affect up to 1% of the population. The pattern of inheritance in the 40% of cases is the same as autosomal dominant Alport syndrome: heterozygous mutations in these genes. The aim of this study is to detect new pathogenic mutations in the COL4A4 gene in the patients previously diagnosed with autosomal Alport syndrome and thin basement membrane nephropathy in our hospital. Methods: We conducted a clinical and genetic study in eleven patients belonging to six unrelated families with aforementioned clinical symptoms and a negative study of COL4A3 gene. The molecular study was made by conformation of sensitive gel electrophoresis (CSGE) and direct sequencing of the fragments that show an altered electrophoretic migration pattern. Results: We found two pathogenic mutations, not yet described: IVS3 + 1G > C is a replacement of Guanine to Cytosine in position +1 of intron 3, in the splicing region, which leads to a pathogenic mutation. c.4267C > T; p.P1423S is a missense mutation, also considered pathogenic. We also found seven new polymorphisms. Conclusions: We describe two new pathogenic mutations, responsible for autosomal dominant Alport syndrome. The other families of the study were undiagnosed owing to problems in the method employed and the possibility of mutations in other genes, giving rise to other diseases with similar symptoms. PMID:25755845

  2. Extensive Analysis of GmFTL and GmCOL Expression in Northern Soybean Cultivars in Field Conditions

    PubMed Central

    Zhu, Jinlong; Lu, Mingyang; Chen, Fulu; Liu, Linpo; Xi, Zhang-Ying; Bachmair, Andreas; Chen, Qingshan; Fu, Yong-Fu

    2015-01-01

    The FLOWERING LOCUS T (FT) gene is a highly conserved florigen gene among flowering plants. Soybean genome encodes six homologs of FT, which display flowering activity in Arabidopsis thaliana. However, their contributions to flowering time in different soybean cultivars, especially in field conditions, are unclear. We employed six soybean cultivars with different maturities to extensively investigate expression patterns of GmFTLs (Glycine max FT-like) and GmCOLs (Glycine max CO-like) in the field conditions. The results show that GmFTL3 is an FT homolog with the highest transcript abundance in soybean, but other GmFTLs may also contribute to flower induction with different extents, because they have more or less similar expression patterns in developmental-, leaf-, and circadian-specific modes. And four GmCOL genes (GmCOL1/2/5/13) may confer to the expression of GmFTL genes. Artificial manipulation of GmFTL expression by transgenic strategy (overexpression and RNAi) results in a distinct change in soybean flowering time, indicating that GmFTLs not only impact on the control of flowering time, but have potential applications in the manipulation of photoperiodic adaptation in soybean. Additionally, transgenic plants show that GmFTLs play a role in formation of the first flowers and in vegetative growth. PMID:26371882

  3. Dominant mutations in the type II collagen gene, COL2A1, produce spondyloepimetaphyseal dysplasia, Strudwick type.

    PubMed

    Tiller, G E; Polumbo, P A; Weis, M A; Bogaert, R; Lachman, R S; Cohn, D H; Rimoin, D L; Eyre, D R

    1995-09-01

    The chondrodysplasias are a heterogeneous group of disorders characterized by abnormal growth or development of cartilage. Current classification is based on mode of inheritance as well as clinical, histologic, and/or radiographic features. A clinical spectrum of chondrodysplasia phenotypes, ranging from mild to perinatal lethal, is due to defects in the gene for type II collagen, COL2A1. This spectrum includes Stickler syndrome, Kniest dysplasia, spondyloepiphyseal dysplasia congenita (SEDC), achondrogenesis type II, and hypochondrogenesis. Individuals affected with these disorders exhibit abnormalities of the growth plate, nucleus pulposus, and vitreous humor, which are tissues that contain type II collagen. The Strudwick type of spondyloepimetaphyseal dysplasia (SEMD) is characterized by disproportionate short stature, pectus carinatum, and scoliosis, as well as dappled metaphyses (which are not seen in SEDC). The phenotype was first described by Murdoch and Walker in 1969, and a series of 14 patients was later reported by Anderson et al. The observation of two affected sibs born to unaffected parents led to the classification of SEMD Strudwick as an autosomal recessive disorder. We now describe the biochemical characterization of defects in alpha 1(II) collagen in three unrelated individuals with SEMD Strudwick, each of which is due to heterozygosity for a unique mutation in COL2A1. Our data support the hypothesis that some cases, if not all cases, of this distinctive chondrodysplasia result from dominant mutations in COL2A1, thus expanding the clinical spectrum of phenotypes associated with this gene. PMID:7550321

  4. Genetic and Epigenetic Factors at COL2A1 and ABCA4 Influence Clinical Outcome in Congenital Toxoplasmosis

    PubMed Central

    Jamieson, Sarra E.; de Roubaix, Lee-Anne; Cortina-Borja, Mario; Tan, Hooi Kuan; Mui, Ernest J.; Cordell, Heather J.; Kirisits, Michael J.; Miller, E. Nancy; Peacock, Christopher S.; Hargrave, Aubrey C.; Coyne, Jessica J.; Boyer, Kenneth; Bessieres, Marie-Hélène; Buffolano, Wilma; Ferret, Nicole; Franck, Jacqueline; Kieffer, François; Meier, Paul; Nowakowska, Dorota E.; Paul, Malgorzata; Peyron, François; Stray-Pedersen, Babill; Prusa, Andrea-Romana; Thulliez, Philippe; Wallon, Martine; Petersen, Eskild; McLeod, Rima; Gilbert, Ruth E.; Blackwell, Jenefer M.

    2008-01-01

    Background Primary Toxoplasma gondii infection during pregnancy can be transmitted to the fetus. At birth, infected infants may have intracranial calcification, hydrocephalus, and retinochoroiditis, and new ocular lesions can occur at any age after birth. Not all children who acquire infection in utero develop these clinical signs of disease. Whilst severity of disease is influenced by trimester in which infection is acquired by the mother, other factors including genetic predisposition may contribute. Methods and Findings In 457 mother-child pairs from Europe, and 149 child/parent trios from North America, we show that ocular and brain disease in congenital toxoplasmosis associate with polymorphisms in ABCA4 encoding ATP-binding cassette transporter, subfamily A, member 4. Polymorphisms at COL2A1 encoding type II collagen associate only with ocular disease. Both loci showed unusual inheritance patterns for the disease allele when comparing outcomes in heterozygous affected children with outcomes in affected children of heterozygous mothers. Modeling suggested either an effect of mother's genotype, or parent-of-origin effects. Experimental studies showed that both ABCA4 and COL2A1 show isoform-specific epigenetic modifications consistent with imprinting. Conclusions These associations between clinical outcomes of congenital toxoplasmosis and polymorphisms at ABCA4 and COL2A1 provide novel insight into the molecular pathways that can be affected by congenital infection with this parasite. PMID:18523590

  5. CNV Analysis in Tourette Syndrome Implicates Large Genomic Rearrangements in COL8A1 and NRXN1

    PubMed Central

    Nag, Abhishek; Bochukova, Elena G.; Kremeyer, Barbara; Campbell, Desmond D.; Muller, Heike; Valencia-Duarte, Ana V.; Cardona, Julio; Rivas, Isabel C.; Mesa, Sandra C.; Cuartas, Mauricio; Garcia, Jharley; Bedoya, Gabriel; Cornejo, William; Herrera, Luis D.; Romero, Roxana; Fournier, Eduardo; Reus, Victor I.; Lowe, Thomas L.; Farooqi, I. Sadaf; Mathews, Carol A.; McGrath, Lauren M.; Yu, Dongmei; Cook, Ed; Wang, Kai; Scharf, Jeremiah M.; Pauls, David L.; Freimer, Nelson B.; Plagnol, Vincent; Ruiz-Linares, Andrés

    2013-01-01

    Tourette syndrome (TS) is a neuropsychiatric disorder with a strong genetic component. However, the genetic architecture of TS remains uncertain. Copy number variation (CNV) has been shown to contribute to the genetic make-up of several neurodevelopmental conditions, including schizophrenia and autism. Here we describe CNV calls using SNP chip genotype data from an initial sample of 210 TS cases and 285 controls ascertained in two Latin American populations. After extensive quality control, we found that cases (N = 179) have a significant excess (P = 0.006) of large CNV (>500 kb) calls compared to controls (N = 234). Amongst 24 large CNVs seen only in the cases, we observed four duplications of the COL8A1 gene region. We also found two cases with ∼400kb deletions involving NRXN1, a gene previously implicated in neurodevelopmental disorders, including TS. Follow-up using multiplex ligation-dependent probe amplification (and including 53 more TS cases) validated the CNV calls and identified additional patients with rearrangements in COL8A1 and NRXN1, but none in controls. Examination of available parents indicates that two out of three NRXN1 deletions detected in the TS cases are de-novo mutations. Our results are consistent with the proposal that rare CNVs play a role in TS aetiology and suggest a possible role for rearrangements in the COL8A1 and NRXN1 gene regions. PMID:23533600

  6. Gene Editing for the Efficient Correction of a Recurrent COL7A1 Mutation in Recessive Dystrophic Epidermolysis Bullosa Keratinocytes.

    PubMed

    Chamorro, Cristina; Mencía, Angeles; Almarza, David; Duarte, Blanca; Büning, Hildegard; Sallach, Jessica; Hausser, Ingrid; Del Río, Marcela; Larcher, Fernando; Murillas, Rodolfo

    2016-01-01

    Clonal gene therapy protocols based on the precise manipulation of epidermal stem cells require highly efficient gene-editing molecular tools. We have combined adeno-associated virus (AAV)-mediated delivery of donor template DNA with transcription activator-like nucleases (TALE) expressed by adenoviral vectors to address the correction of the c.6527insC mutation in the COL7A1 gene, causing recessive dystrophic epidermolysis bullosa in a high percentage of Spanish patients. After transduction with these viral vectors, high frequencies of homology-directed repair were found in clones of keratinocytes derived from a recessive dystrophic epidermolysis bullosa (RDEB) patient homozygous for the c.6527insC mutation. Gene-edited clones recovered the expression of the COL7A1 transcript and collagen VII protein at physiological levels. In addition, treatment of patient keratinocytes with TALE nucleases in the absence of a donor template DNA resulted in nonhomologous end joining (NHEJ)-mediated indel generation in the vicinity of the c.6527insC mutation site in a large proportion of keratinocyte clones. A subset of these indels restored the reading frame of COL7A1 and resulted in abundant, supraphysiological expression levels of mutant or truncated collagen VII protein. Keratinocyte clones corrected both by homology-directed repair (HDR) or NHEJ were used to regenerate skin displaying collagen VII in the dermo-epidermal junction. PMID:27045209

  7. A novel collagen/platelet-rich plasma (COL/PRP) scaffold: preparation and growth factor release analysis.

    PubMed

    Zhang, Xiujie; Wang, Jingwei; Ren, Mingguang; Li, Lifeng; Wang, Qingwen; Hou, Xiaohua

    2016-06-01

    Platelet-rich plasma (PRP) has been widely used in clinical practice for more than 20 years because it causes the release of many growth factors. However, the burst release pattern and short release period of PRP have become obstacles to its application. An optimal controllable release system is an urgent need for researchers. This study investigated whether collagen/PRP (COL/PRP) scaffolds can serve as a vehicle for the controllable release of growth factors. We fabricated a novel scaffold that integrates PRP activated by thrombin or collagen into type I collagen. The mechanical properties, cytotoxicity, and transforming growth factor β1 (TGF-β1), platelet derived growth factor (PDGF), fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) content were evaluated. Our results demonstrate that the COL/PRP scaffolds were not cytotoxic to L-929 fibroblasts. The PDGF and FGF content in the thrombin group was at a higher level and lasted for a long period of time. Collagen and thrombin played the same role in the release of TGF-β1 and VEGF. These data suggest that the novel COL/PRP scaffolds provide a carrier for the controllable release of growth factors and may be used in tissue- regenerative therapies. PMID:26951554

  8. THE SOLAR NEIGHBORHOOD. XXVI. AP Col: THE CLOSEST (8.4 pc) PRE-MAIN-SEQUENCE STAR

    SciTech Connect

    Riedel, Adric R.; Henry, Todd J.; Jao, Wei-Chun; Murphy, Simon J.; Melis, Carl; Subasavage, John P. E-mail: thenry@chara.gsu.edu E-mail: murphysj@mso.anu.edu.au E-mail: jsubasavage@ctio.noao.edu

    2011-10-15

    We present the results of a multi-technique investigation of the M4.5Ve flare star AP Col, which we discover to be the nearest pre-main-sequence star. These include astrometric data from the CTIO 0.9 m, from which we derive a proper motion of 342.0 {+-} 0.5 mas yr{sup -1}, a trigonometric parallax of 119.21 {+-} 0.98 mas (8.39 {+-} 0.07 pc), and photometry and photometric variability at optical wavelengths. We also provide spectroscopic data, including radial velocity (22.4 {+-} 0.3 km s{sup -1}), lithium equivalent width (EW) (0.28 {+-} 0.02 A), H{alpha} EW (-6.0 to -35 A), vsin i (11 {+-} 1 km s{sup -1}), and gravity indicators from the Siding Spring 2.3 m WiFeS, Lick 3 m Hamilton echelle, and Keck-I HIRES echelle spectrographs. The combined observations demonstrate that AP Col is the closer of only two known systems within 10 pc of the Sun younger than 100 Myr. Given its space motion and apparent age of 12-50 Myr, AP Col is likely a member of the recently proposed {approx}40 Myr old Argus/IC 2391 Association.

  9. Dysspondyloenchondromatosis (DSC) associated with COL2A1 mutation: Clinical and radiological overlap with spondyloepimetaphyseal dysplasia-Strudwick type (SEMD-S).

    PubMed

    Merrick, Blair; Calder, Alistair; Wakeling, Emma

    2015-12-01

    Dysspondyloenchondromatosis (DSC) is a rare skeletal dysplasia characterized by enchondroma-like lesions and anisospondyly. The former leads to discrepancies in limb length, and the latter, to progressive kyphoscoliosis. Two recent cases have highlighted the genetic heterogeneity of DSC, one demonstrating the presence and, the other, the absence of a COL2A1 mutation. This may have important clinical implications, for example, screening for complications including atlanto-axial instability associated with type II collagenopathies, as well as long-term patient management. We report on a case with radiographic features of DSC with overlap into the type II collagenopathy spondyloepimetaphyseal dysplasia, Strudwick type, who was found to carry a novel heterozygous mutation in the COL2A1 gene. Testing for COL2A1 mutations should be performed in all patients with radiological features of DSC. Further research is needed to identify the underlying molecular cause in cases where no COL2A1 mutation is identified. PMID:26250472

  10. Identification of the collagen type 1 alpha 1 gene (COL1A1) as a candidate survival-related factor associated with hepatocellular carcinoma

    PubMed Central

    2014-01-01

    Background Hepatocellular carcinoma (HCC) is one of the major causes of cancer-related death especially among Asian and African populations. It is urgent that we identify carcinogenesis-related genes to establish an innovative treatment strategy for this disease. Methods Triple-combination array analysis was performed using one pair each of HCC and noncancerous liver samples from a 68-year-old woman. This analysis consists of expression array, single nucleotide polymorphism array and methylation array. The gene encoding collagen type 1 alpha 1 (COL1A1) was identified and verified using HCC cell lines and 48 tissues from patients with primary HCC. Results Expression array revealed that COL1A1 gene expression was markedly decreased in tumor tissues (log2 ratio –1.1). The single nucleotide polymorphism array showed no chromosomal deletion in the locus of COL1A1. Importantly, the methylation value in the tumor tissue was higher (0.557) than that of the adjacent liver tissue (0.008). We verified that expression of this gene was suppressed by promoter methylation. Reactivation of COL1A1 expression by 5-aza-2′-deoxycytidine treatment was seen in HCC cell lines, and sequence analysis identified methylated CpG sites in the COL1A1 promoter region. Among 48 pairs of surgical specimens, 13 (27.1%) showed decreased COL1A1 mRNA expression in tumor sites. Among these 13 cases, 10 had promoter methylation at the tumor site. The log-rank test indicated that mRNA down-regulated tumors were significantly correlated with a poor overall survival rate (P = 0.013). Conclusions Triple-combination array analysis successfully identified COL1A1 as a candidate survival-related gene in HCCs. Epigenetic down-regulation of COL1A1 mRNA expression might have a role as a prognostic biomarker of HCC. PMID:24552139

  11. COL1A1 and miR-29b show lower expression levels during osteoblast differentiation of bone marrow stromal cells from Osteogenesis Imperfecta patients

    PubMed Central

    2014-01-01

    Background The majority of Osteogenesis Imperfecta (OI) cases are caused by mutations in one of the two genes, COL1A1 and COL1A2 encoding for the two chains that trimerize to form the procollagen 1 molecule. However, alterations in gene expression and microRNAs (miRNAs) are responsible for the regulation of cell fate determination and may be evolved in OI phenotype. Methods In this work, we analyzed the coding region and intron/exon boundaries of COL1A1 and COL1A2 genes by sequence analysis using an ABI PRISM 3130 automated sequencer and Big Dye Terminator Sequencing protocol. COL1A1 and miR-29b expression were also evaluated during the osteoblastic differentiation of mesenchymal stem cell (MSC) by qRT-PCR using an ABI7500 Sequence Detection System. Results We have identified eight novel mutations, where of four may be responsible for OI phenotype. COL1A1 and miR-29b showed lower expression values in OI type I and type III samples. Interestingly, one type III OI sample from a patient with Bruck Syndrome showed COL1A1 and miR-29b expressions alike those from normal samples. Conclusions Results suggest that the miR-29b mechanism directed to regulate collagen protein accumulation during mineralization is dependent upon the amount of COL1A1 mRNA. Taken together, results indicate that the lower levels observed in OI samples were not sufficient for the induction of miR-29b. PMID:24767406

  12. Progression of Alport Kidney Disease in Col4a3 Knock Out Mice Is Independent of Sex or Macrophage Depletion by Clodronate Treatment

    PubMed Central

    Kim, Munkyung; Piaia, Alessandro; Shenoy, Neeta; Kagan, David; Gapp, Berangere; Kueng, Benjamin; Weber, Delphine; Dietrich, William; Ksiazek, Iwona

    2015-01-01

    Alport syndrome is a genetic disease of collagen IV (α3, 4, 5) resulting in renal failure. This study was designed to investigate sex-phenotype correlations and evaluate the contribution of macrophage infiltration to disease progression using Col4a3 knock out (Col4a3KO) mice, an established genetic model of autosomal recessive Alport syndrome. No sex differences in the evolution of body mass loss, renal pathology, biomarkers of tubular damage KIM-1 and NGAL, or deterioration of kidney function were observed during the life span of Col4a3KO mice. These findings confirm that, similar to human autosomal recessive Alport syndrome, female and male Col4a3KO mice develop renal failure at the same age and with similar severity. The specific contribution of macrophage infiltration to Alport disease, one of the prominent features of the disease in human and Col4a3KO mice, remains unknown. This study shows that depletion of kidney macrophages in Col4a3KO male mice by administration of clodronate liposomes, prior to clinical onset of disease and throughout the study period, does not protect the mice from renal failure and interstitial fibrosis, nor delay disease progression. These results suggest that therapy targeting macrophage recruitment to kidney is unlikely to be effective as treatment of Alport syndrome. PMID:26555339

  13. Physical and linkage mapping of the human and murine genes for the [alpha]1 chain of type IX collagen (COL9A1)

    SciTech Connect

    Warman, M.L. Children's Hospital Tiller, G.E.; Polumbo, P.A. ); Seldin, M.F.; Rochelle, J.M. ); Knoll, J.H.M.; Cheng, Sou De ); Olsen, B.R. )

    1993-09-01

    The IX collagen, a member of the FACIT family of extracellular matrix proteins, is a heterotrimer composed of three genetically distinct [alpha] chains. The cDNAs for the human and mouse [alpha]1(IX) chains have been cloned. In this paper the authors confirm the mapping of the human COL9A1 gene to chromosome 6q12-q13 by fluorescence in situ hybridization utilizing two genomic clones which also contain short tandem repeat polymorphisms. They also report the characterization of these repeats and their incorporation into the chromosome 6 linkage map. The COL9A1 locus shows no recombination with the marker D6Z1 (Z = 27.61 at [theta] = 0) and identifies the most likely locus order of KRAS1P-[D6Z1-COL9A1]-D6S30. In addition, using an interspecific backcross panel, they have mapped murine Col9a1 to mouse chromosome 1. Together with other comparative mapping results, these data suggest that the pericentric region of human chromosome 6 is homologous to the most proximal segment of mouse chromosome 1. These data may facilitate linkage studies with COL9A1 (or col9a1) as a candidate gene for hereditary chondrodysplasias and osteoarthritis. 35 refs., 2 figs., 2 tabs.

  14. Ledge-type Co/L10-FePt exchange-coupled composites

    NASA Astrophysics Data System (ADS)

    Speliotis, Th.; Giannopoulos, G.; Niarchos, D.; Li, W. F.; Hadjipanayis, G.; Barucca, G.; Agostinelli, E.; Laureti, S.; Peddis, D.; Testa, A. M.; Varvaro, G.

    2016-06-01

    FePt-based exchange-coupled composites consisting of a magnetically hard L10-FePt phase exchange-coupled with a soft ferromagnetic material are promising candidates for future ultra-high density (>1 Tbit/in2) perpendicular magnetic recording media, also being of interest for other applications including spin torque oscillators and micro-electro-mechanical systems, among others. In this paper, the effect of the thickness of a soft Co layer (3 < thCo < 20 nm) on the magnetic behavior of ledge-type fcc(100)-Co/L10(001)-FePt composites deposited on an MgO (100) substrate is systematically studied by combining morpho-structural analyses and angular magnetization measurements. Starting from a film consisting of isolated L10(001)-FePt islands, the ledge-type structure was obtained by depositing a Co layer that either covered the FePt islands or filled-up the inter-island region, gradually forming a continuous layer with increasing Co thickness. A perpendicular anisotropy was maintained up to thCo ˜ 9.5 nm and a significant reduction in the coercivity (about 50% for thCo ˜ 3 nm) with the increase in thCo was observed, indicating that, by coupling hard FePt and soft Co phases in a ledge-type configuration, the writability can be greatly improved. Recoil loops' measurements confirmed the exchange-coupled behavior, reinforcing a potential interest in these systems for future magnetic recording media.

  15. Molecular Evolution and Phylogenetic Analysis of Eight COL Superfamily Genes in Group I Related to Photoperiodic Regulation of Flowering Time in Wild and Domesticated Cotton (Gossypium) Species

    PubMed Central

    Zhang, Rui; Ding, Jian; Liu, Chunxiao; Cai, Caiping; Zhou, Baoliang; Zhang, Tianzhen; Guo, Wangzhen

    2015-01-01

    Flowering time is an important ecological trait that determines the transition from vegetative to reproductive growth. Flowering time in cotton is controlled by short-day photoperiods, with strict photoperiod sensitivity. As the CO-FT (CONSTANS-FLOWER LOCUS T) module regulates photoperiodic flowering in several plants, we selected eight CONSTANS genes (COL) in group I to detect their expression patterns in long-day and short-day conditions. Further, we individually cloned and sequenced their homologs from 25 different cotton accessions and one outgroup. Finally, we studied their structures, phylogenetic relationship, and molecular evolution in both coding region and three characteristic domains. All the eight COLs in group I show diurnal expression. In the orthologous and homeologous loci, each gene structure in different cotton species is highly conserved, while length variation has occurred due to insertions/deletions in intron and/or exon regions. Six genes, COL2 to COL5, COL7 and COL8, exhibit higher nucleotide diversity in the D-subgenome than in the A-subgenome. The Ks values of 98.37% in all allotetraploid cotton species examined were higher in the A-D and At-Dt comparison than in the A-At and D-Dt comparisons, and the Pearson’s correlation coefficient (r) of Ks between A vs. D and At vs. Dt also showed positive, high correlations, with a correlation coefficient of at least 0.797. The nucleotide polymorphism in wild species is significantly higher compared to G. hirsutum and G. barbadense, indicating a genetic bottleneck associated with the domesticated cotton species. Three characteristic domains in eight COLs exhibit different evolutionary rates, with the CCT domain highly conserved, while the B-box and Var domain much more variable in allotetraploid species. Taken together, COL1, COL2 and COL8 endured greater selective pressures during the domestication process. The study improves our understanding of the domestication-related genes/traits during cotton

  16. Biochemical and functional studies of ColTx-I, a new myotoxic phospholipase A2 isolated from Crotalus oreganus lutosus (Great Basin rattlesnake) snake venom.

    PubMed

    Almeida, J R; Resende, L M; Silva, A G; Ribeiro, R I M A; Stábeli, R G; Soares, A M; Calderon, L A; Marangoni, S; Da Silva, S L

    2016-07-01

    Commonly, phospholipases A2 (PLA2s) play key roles in the pathogenesis of the local tissue damage characteristic of crotaline and viperine snake envenomations. Crotalus oreganus lutosus snake venom has not been extensively studied; therefore, the characterization of its components represents a valuable biotechnological tool for studying pathophysiological processes of envenoming and for gaining a deeper understanding of its biological effects. In this study, for the first time, a basic PLA2 myotoxin, ColTx-I, was purified from C. o. lutosus through two chromatographic steps. ColTx-I is monomeric with calculated molecular mass weight (Mw) of 14,145 Da and a primary structure closely related to basic PLA2s from viperid venoms. The pure enzyme has a specific activity of 15.87 ± 0.65 nmol/min/mg at optimal conditions (pH 8.0 and 37 °C). ColTx-I activity was found to be dependent on Ca(2+), as its substitution by other ionic species as well as the addition of chelating agents significantly reduced its phospholipase activity. In vivo, ColTx-I triggered dose-dependent inflammatory responses, measured using the paw edema model, with an increase in IL-6 levels, systemic and local myotoxicity, characterized by elevated plasma creatine kinase activity. ColTx-I induced a complex series of degenerative events associated with edema, inflammatory infiltrate and skeletal muscle necrosis. These biochemical and functional results suggest that ColTx-I, a myotoxic and inflammatory mediator, plays a relevant role in C. o. lutosus envenomation. Thus, detailed studies on its mechanism of action, such as evaluating the synergism between ColTx-I and other venom components may reveal targets for the development of more specific and effective therapies. PMID:26996495

  17. ER stress and basement membrane defects combine to cause glomerular and tubular renal disease resulting from Col4a1 mutations in mice

    PubMed Central

    Jones, Frances E.; Bailey, Matthew A.; Murray, Lydia S.; Lu, Yinhui; McNeilly, Sarah; Schlötzer-Schrehardt, Ursula; Lennon, Rachel; Sado, Yoshikazu; Brownstein, David G.; Mullins, John J.; Kadler, Karl E.; Van Agtmael, Tom

    2016-01-01

    ABSTRACT Collagen IV is a major component of basement membranes, and mutations in COL4A1, which encodes collagen IV alpha chain 1, cause a multisystemic disease encompassing cerebrovascular, eye and kidney defects. However, COL4A1 renal disease remains poorly characterized and its pathomolecular mechanisms are unknown. We show that Col4a1 mutations in mice cause hypotension and renal disease, including proteinuria and defects in Bowman's capsule and the glomerular basement membrane, indicating a role for Col4a1 in glomerular filtration. Impaired sodium reabsorption in the loop of Henle and distal nephron despite elevated aldosterone levels indicates that tubular defects contribute to the hypotension, highlighting a novel role for the basement membrane in vascular homeostasis by modulation of the tubular response to aldosterone. Col4a1 mutations also cause diabetes insipidus, whereby the tubular defects lead to polyuria associated with medullary atrophy and a subsequent reduction in the ability to upregulate aquaporin 2 and concentrate urine. Moreover, haematuria, haemorrhage and vascular basement membrane defects confirm an important vascular component. Interestingly, although structural and compositional basement membrane defects occurred in the glomerulus and Bowman's capsule, no tubular basement membrane defects were detected. By contrast, medullary atrophy was associated with chronic ER stress, providing evidence for cell-type-dependent molecular mechanisms of Col4a1 mutations. These data show that both basement membrane defects and ER stress contribute to Col4a1 renal disease, which has important implications for the development of treatment strategies for collagenopathies. PMID:26839400

  18. Evolution of a highly vulnerable ice-cored moraine: Col des Gentianes, Swiss Alps

    NASA Astrophysics Data System (ADS)

    Ravanel, L.; Lambiel, C.; Oppikofer, T.; Mazotti, B.; Jaboyedoff, M.

    2012-04-01

    Rock mass movements are dominant in the morphodynamics of high mountain rock slopes and are at the origin of significant risks for people who attend these areas and for infrastructures that are built on (mountain huts, cable cars, etc.). These risks are becoming greater because of permafrost degradation and glacier retreat, two consequences of the global warming. These two commonly associated factors may affect slope stability by changing mechanical properties of the interstitial ice and modifying the mechanical constraints in these rock slopes. Between 1977 and 1979, significant works were carried out on the Little Ice Age moraine of the Tortin glacier at the Col des Gentianes (2894 m), in the Mont Fort area (Verbier, Switzerland), for the construction of a cable car station and a restaurant. Since the early 1980s, the glacier drastically retreated and the moraine became unstable: its inner slope has retreated for several meters. Various observations and geoelectric measurements indicate that significant volume of massive ice mass is still present within the moraine (ice-cored moraine). Its melting could therefore increase the instability of the moraine. Since 2007, the moraine is surveyed by terrestrial laser scanning (TLS) in order to characterize its evolution: 8 campaigns were conducted between July 2007 and October 2011. The comparison of the high resolution 3D models so obtained allowed the detection and quantification of mass movements that have affected the moraine over this period, essentially by calculating difference maps (shortest oblique distances between two models). Between July 2007 and October 2011, 7 landslides were measured, involving volumes between 87 and 1138 m3. The most important of these occurred during the summers 2009 and 2011. TLS data also allowed identifying: (i) two main areas affected by slower but sometimes substantial movements (displacements of blocks on more than 2 m during a summer period); (ii) significant deposits of

  19. Variable Bone Fragility Associated With an Amish COL1A2 Variant and a Knock-in Mouse Model

    PubMed Central

    Daley, Ethan; Streeten, Elizabeth A; Sorkin, John D; Kuznetsova, Natalia; Shapses, Sue A; Carleton, Stephanie M; Shuldiner, Alan R; Marini, Joan C; Phillips, Charlotte L; Goldstein, Steven A; Leikin, Sergey; McBride, Daniel J

    2010-01-01

    Osteogenesis imperfecta (OI) is a heritable form of bone fragility typically associated with a dominant COL1A1 or COL1A2 mutation. Variable phenotype for OI patients with identical collagen mutations is well established, but phenotype variability is described using the qualitative Sillence classification. Patterning a new OI mouse model on a specific collagen mutation therefore has been hindered by the absence of an appropriate kindred with extensive quantitative phenotype data. We benefited from the large sibships of the Old Order Amish (OOA) to define a wide range of OI phenotypes in 64 individuals with the identical COL1A2 mutation. Stratification of carrier spine (L1–4) areal bone mineral density (aBMD) Z-scores demonstrated that 73% had moderate to severe disease (less than −2), 23% had mild disease (−1 to −2), and 4% were in the unaffected range (greater than −1). A line of knock-in mice was patterned on the OOA mutation. Bone phenotype was evaluated in four F1 lines of knock-in mice that each shared approximately 50% of their genetic background. Consistent with the human pedigree, these mice had reduced body mass, aBMD, and bone strength. Whole-bone fracture susceptibility was influenced by individual genomic factors that were reflected in size, shape, and possibly bone metabolic regulation. The results indicate that the G610C OI (Amish) knock-in mouse is a novel translational model to identify modifying genes that influence phenotype and for testing potential therapies for OI. © 2010 American Society for Bone and Mineral Research PMID:19594296

  20. Transcriptional Repression of the Dspp Gene Leads to Dentinogenesis Imperfecta Phenotype in Col1a1-Trps1 Transgenic Mice

    PubMed Central

    Napierala, Dobrawa; Sun, Yao; Maciejewska, Izabela; Bertin, Terry K; Dawson, Brian; D'Souza, Rena; Qin, Chunlin; Lee, Brendan

    2012-01-01

    Dentinogenesis imperfecta (DGI) is a hereditary defect of dentin, a calcified tissue that is the most abundant component of teeth. Most commonly, DGI is manifested as a part of osteogenesis imperfecta (OI) or the phenotype is restricted to dental findings only. In the latter case, DGI is caused by mutations in the DSPP gene, which codes for dentin sialoprotein (DSP) and dentin phosphoprotein (DPP). Although these two proteins together constitute the majority of noncollagenous proteins of the dentin, little is known about their transcriptional regulation. Here we demonstrate that mice overexpressing the Trps1 transcription factor (Col1a1-Trps1 mice) in dentin-producing cells, odontoblasts, present with severe defects of dentin formation that resemble DGI. Combined micro–computed tomography (µCT) and histological analyses revealed tooth fragility due to severe hypomineralization of dentin and a diminished dentin layer with irregular mineralization in Col1a1-Trps1 mice. Biochemical analyses of noncollagenous dentin matrix proteins demonstrated decreased levels of both DSP and DPP proteins in Col1a1-Trps1 mice. On the molecular level, we demonstrated that sustained high levels of Trps1 in odontoblasts lead to dramatic decrease of Dspp expression as a result of direct inhibition of the Dspp promoter by Trps1. During tooth development Trps1 is highly expressed in preodontoblasts, but in mature odontoblasts secreting matrix its expression significantly decreases, which suggests a Trps1 role in odontoblast development. In these studies we identified Trps1 as a potent inhibitor of Dspp expression and the subsequent mineralization of dentin. Thus, we provide novel insights into mechanisms of transcriptional dysregulation that leads to DGI. © 2012 American Society for Bone and Mineral Research. PMID:22508542

  1. Identification of COL3A1 and RAB2A as novel translocation partner genes of PLAG1 in lipoblastoma.

    PubMed

    Yoshida, Hideki; Miyachi, Mitsuru; Ouchi, Kazutaka; Kuwahara, Yasumichi; Tsuchiya, Kunihiko; Iehara, Tomoko; Konishi, Eiichi; Yanagisawa, Akio; Hosoi, Hajime

    2014-07-01

    Lipoblastoma is a rapidly growing, benign neoplasm in children. Surgical excision is usually curative, with a recurrence rate of about 20%. Because the histology of lipoblastoma is heterogeneous and overlaps with other lipomatous tumors, some lipoblastoma cases have been difficult to diagnose. The detection of PLAG1 gene rearrangement is useful for the diagnosis of lipoblastoma. Three fusion partner genes are known in relation to PLAG1 in lipoblastoma HAS2 at 8q24.1, COL1A2 at 7q22, and RAD51L1 at 14q24. Herein, we describe another two novel fusion genes in lipoblastoma tumor specimens. We checked six tumors for the presence of two known fusion genes, HAS2-PLAG1 and COL1A2-PLAG1. Only HAS2-PLAG1 was found in one of the cases. Next, we attempted to identify potential PLAG1 fusion partners using 5'RACE. Sequence analysis revealed two novel fusion genes, COL3A1-PLAG1 in three cases and RAB2A-PLAG1 in one case, respectively. As a result of the translocations, the constitutively active promoter of the partner gene drives the ectopic expression of PLAG1. We also evaluated whether a high level of PLAG1 expression can be used to help differentiate lipomatous tumors. PLAG1 expression was evaluated by real-time PCR in five lipoblastoma tumor specimens. The expressions were 70-150 times higher in lipoblastomas than in human adipocytes. However, PLAG1 expression was low in one case of lipoma. These results demonstrate that PLAG1 overexpression is a potential marker of lipoblastoma. Our findings, in agreement with previous studies, show that lipoblastoma is a group of lipomatous tumors with PLAG1 rearrangement and overexpression. © 2014 Wiley Periodicals, Inc. PMID:24700772

  2. Overrepresentation of the COL3A1 AA genotype in Polish skiers with anterior cruciate ligament injury.

    PubMed

    Stępień-Słodkowska, M; Ficek, K; Maciejewska-Karłowska, A; Sawczuk, M; Ziętek, P; Król, P; Zmijewski, P; Pokrywka, A; Cięszczyk, P

    2015-06-01

    Although various intrinsic and extrinsic risk factors for anterior cruciate ligament (ACL) rupture have been identified, the exact aetiology of the injury is not yet fully understood. Type III collagen is an important factor in the repair of connective tissue, and certain gene polymorphisms may impair the tensile strength. The aim of this study was to examine the association of the COL3A1 rs1800255 polymorphism with ACL rupture in Polish male recreational skiers. A total of 321 male Polish recreational skiers were recruited for this study; 138 had surgically diagnosed primary ACL ruptures (ACL-injured group) and 183 were apparently healthy male skiers (control group - CON) who had no self-reported history of ligament or tendon injury. Both groups had a comparable level of exposure to ACL injury. Genomic DNA was extracted from the oral epithelial cells. All samples were genotyped on a real-time polymerase chain reaction instrument. The genotype distribution in the ACL-injured group was significantly different than in CON (respectively: AA=10.1 vs 2.2%, AG=22.5 vs 36.1, GG=67.4 vs 61.8%; p=0.0087). The AA vs AG+GG genotype of COL3A1 (odds ratio (OR)=5.05; 95% confidence interval (CI), 1.62-15.71, p=0.003) was significantly overrepresented in the ACL-injured group compared with CON. The frequency of the A allele was higher in the ACL-injured group (21.4%) compared with CON (20.2%), but the difference was not statistically significant (p=0.72). This study revealed an association between the COL3A1 rs1800255 polymorphism and ACL ruptures in Polish skiers. PMID:26060338

  3. Homozygosity Mapping and Whole Exome Sequencing Reveal a Novel Homozygous COL18A1 Mutation Causing Knobloch Syndrome

    PubMed Central

    Piri, Niloofar; Nürnberg, Gudrun; Saleh-Gohari, Nasrollah; Haghighi, Amirreza; Neidhardt, John; Nürnberg, Peter; Berger, Wolfgang

    2014-01-01

    The aim of this study was to identify the genetic basis of a chorioretinal dystrophy with high myopia of unknown origin in a child of a consanguineous marriage. The proband and ten family members of Iranian ancestry participated in this study. Linkage analysis was carried out with DNA samples of the proband and her parents by using the Human SNP Array 6.0. Whole exome sequencing (WES) was performed with the patients’ DNA. Specific sequence alterations within the homozygous regions identified by whole exome sequencing were verified by Sanger sequencing. Upon genetic analysis, a novel homozygous frameshift mutation was found in exon 42 of the COL18A1 gene in the patient. Both parents were heterozygous for this sequence variation. Mutations in COL18A1 are known to cause Knobloch syndrome (KS). Retrospective analysis of clinical records of the patient revealed surgical removal of a meningocele present at birth. The clinical features shown by our patient were typical of KS with the exception of chorioretinal degeneration which is a rare manifestation. This is the first case of KS reported in a family of Iranian ancestry. We identified a novel disease-causing (deletion) mutation in the COL18A1 gene leading to a frameshift and premature stop codon in the last exon. The mutation was not present in SNP databases and was also not found in 192 control individuals. Its localization within the endostatin domain implicates a functional relevance of endostatin in KS. A combined approach of linkage analysis and WES led to a rapid identification of the disease-causing mutation even though the clinical description was not completely clear at the beginning. PMID:25392994

  4. Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    PubMed Central

    Maguire, Kim R.; Sidén, Åke; Potácová, Zuzana

    2014-01-01

    Tetracycline analogues (TCNAs) have been shown to inhibit matrix metalloproteinases and to induce apoptosis in several cancer cell types. In the present study, the cytotoxic effects of TCNAs doxycycline (DOXY), minocycline (MINO) and chemically modified tetracycline-3 (COL-3) were investigated in the human acute myeloid leukemia HL-60 cell line. Cells were incubated with TCNAs in final concentrations of 0.5–100 µg/ml for 24 h. Viability of the leukemic cells was inhibited in a concentration-dependent manner using resazurin assay. The estimated IC50s were 9.2 µg/ml for DOXY, 9.9 µg/ml for MINO and 1.3 µg/ml for COL-3. All three TCNAs induced potent cytotoxic effects and cell death. Apoptosis, which was assessed by morphological changes and annexin V positivity, was concentration- and time-dependent following incubation with any one of the drugs. TCNAs induced DNA double strand breaks soon after treatment commenced as detected by γH2AX and western blot. The loss of mitochondrial membrane potential (Δψm), caspase activation and cleavage of PARP and Bcl-2 were observed; however, the sequence of events differed among the drugs. Pancaspase inhibitor Z-VAD-FMK improved survival of TCNAs-treated cells and decreased TCNAs-induced apoptosis. In summary, we demonstrated that TCNAs had a cytotoxic effect on the HL-60 leukemic cell line. Apoptosis was induced via mitochondria-mediated and caspase-dependent pathways in HL-60 cells by all three TCNAs. COL-3 exerted the strongest anti-proliferative and pro-apoptotic effects in concentrations that have been achieved in human plasma in reported clinical trials. These results indicate that there is a therapeutic potential of TCNAs in leukemia. PMID:25502932

  5. Mutations in the COL5A1 gene are causal in the Ehlers-Danlos syndromes I and II.

    PubMed Central

    De Paepe, A; Nuytinck, L; Hausser, I; Anton-Lamprecht, I; Naeyaert, J M

    1997-01-01

    The Ehlers-Danlos syndrome (EDS) is a heterogeneous connective-tissue disorder of which at least nine subtypes are recognized. Considerable clinical overlap exists between the EDS I and II subtypes, suggesting that both are allelic disorders. Recent evidence based on linkage and transgenic mice studies suggest that collagen V is causally involved in human EDS. Collagen V forms heterotypic fibrils with collagen I in many tissues and plays an important role in collagen I fibrillogenesis. We have identified a mutation in COL5A1, the gene encoding the pro(alpha)1(V) collagen chain, segregating with EDS I in a four-generation family. The mutation causes the substitution of the most 5' cysteine residue by a serine within a highly conserved sequence of the pro(alpha)1(V) C-propeptide domain and causes reduction of collagen V by preventing incorporation of the mutant pro(alpha)1(V) chains in the collagen V trimers. In addition, we have detected splicing defects in the COL5A1 gene in a patient with EDS I and in a family with EDS II. These findings confirm the causal role of collagen V in at least a subgroup of EDS I, prove that EDS I and II are allelic conditions, and represent a, so far, unique example of a human collagen disorder caused by substitution of a highly conserved cysteine residue in the C-propeptide domain of a fibrillar collagen. Images Figure 1 Figure 2 Figure 3 Figure 5 PMID:9042913

  6. Metabolic Footprint Analysis Uncovers Strain Specific Overflow Metabolism and D-Isoleucine Production of Staphylococcus Aureus COL and HG001

    PubMed Central

    Dörries, Kirsten; Lalk, Michael

    2013-01-01

    During infection processes, Staphylococcus aureus is able to survive within the host and to invade tissues and cells. For studying the interaction between the pathogenic bacterium and the host cell, the bacterial growth behaviour and its metabolic adaptation to the host cell environment provides first basic information. In the present study, we therefore cultivated S. aureus COL and HG001 in the eukaryotic cell culture medium RPMI 1640 and analyzed the extracellular metabolic uptake and secretion patterns of both commonly used laboratory strains. Extracellular accumulation of D-isoleucine was detected starting during exponential growth of COL and HG001 in RPMI medium. This non-canonical D-amino acid is known to play a regulatory role in adaptation processes. Moreover, individual uptake of glucose, accumulation of acetate, further overflow metabolites, and intermediates of the branched-chain amino acid metabolism constitute unique metabolic footprints. Altogether these time-resolved footprint analyses give first metabolic insights into staphylococcal growth behaviour in a culture medium used for infection related studies. PMID:24312553

  7. Dual reporter transgene driven by 2.3Col1a1 promoter is active in differentiated osteoblasts

    NASA Technical Reports Server (NTRS)

    Marijanovic, Inga; Jiang, Xi; Kronenberg, Mark S.; Stover, Mary Louise; Erceg, Ivana; Lichtler, Alexander C.; Rowe, David W.

    2003-01-01

    AIM: As quantitative and spatial analyses of promoter reporter constructs are not easily performed in intact bone, we designed a reporter gene specific to bone, which could be analyzed both visually and quantitatively by using chloramphenicol acetyltransferase (CAT) and a cyan version of green fluorescent protein (GFPcyan), driven by a 2.3-kb fragment of the rat collagen promoter (Col2.3). METHODS: The construct Col2.3CATiresGFPcyan was used for generating transgenic mice. Quantitative measurement of promoter activity was performed by CAT analysis of different tissues derived from transgenic animals; localization was performed by visualized GFP in frozen bone sections. To assess transgene expression during in vitro differentiation, marrow stromal cell and neonatal calvarial osteoblast cultures were analyzed for CAT and GFP activity. RESULTS: In mice, CAT activity was detected in the calvaria, long bone, teeth, and tendon, whereas histology showed that GFP expression was limited to osteoblasts and osteocytes. In cell culture, increased activity of CAT correlated with increased differentiation, and GFP activity was restricted to mineralized nodules. CONCLUSION: The concept of a dual reporter allows a simultaneous visual and quantitative analysis of transgene activity in bone.

  8. Spectrum of mutations in the COL4A5 collagen gene in X-linked Alport syndrome.

    PubMed Central

    Knebelmann, B.; Breillat, C.; Forestier, L.; Arrondel, C.; Jacassier, D.; Giatras, I.; Drouot, L.; Deschênes, G.; Grünfeld, J. P.; Broyer, M.; Gubler, M. C.; Antignac, C.

    1996-01-01

    Alport syndrome is a mainly X-linked hereditary disease of basement membranes that is characterized by progressive renal failure, deafness, and ocular lesions. It is associated with mutations of the COL4A5 gene located at Xq22 and encoding the alpha5 chain of type IV collagen. We have screened 48 of the 51 exons of the COL4A5 gene by SSCP analysis and have identified 64 mutations and 10 sequence variants among 131 unrelated Alport syndrome patients. This represents a mutation-detection rate of 50%. There were no hot-spot mutations and no recurrent mutations in our population. The identified mutations were 6 nonsense mutations, 12 frameshift mutations, 17 splice-site mutations, and 29 missense mutations, 27 of the latter being glycine substitutions in the collagenous domain. Two of these occurred on the same allele in one patient and segregated with the disease in the family. We showed that some of the glycine substitutions could be associated with the lack of immunological expression of the alpha3(IV)-alpha5(IV) collagen chains in the glomerular basement membrane. Images Figure 1 Figure 2 PMID:8940267

  9. Proteome analysis differentiates between two highly homologues germin-like proteins in Arabidopsis thaliana ecotypes Col-0 and Ws-2.

    PubMed

    Schlesier, Bernhard; Berna, Anne; Bernier, François; Mock, Hans-Peter

    2004-06-01

    A proteome approach based on 2-D gel electrophoresis (2-DE) was used to compare the protein patterns of the Arabidopsis ecotypes Col-0 and Ws-2. In leaf extracts a pair of protein spots were found to be diagnostic for each of the lines. Both pairs of spots were identified as closely related germin-like proteins differing in only one amino acid by using peptide mass finger printing of tryptic digests and by gaining additional data from post-source decay spectra in the MALDI-TOF analysis. Western blot analysis after separation of protein extracts by 2-DE confirmed results from Coomassie blue-stained gels and revealed additional immunoreactive spots for both ecotypes most likely representing dimers of the spots first identified. Western blot analysis and mass spectrometrical identification of the corresponding weakly stained protein in Coomassie blue-stained gels of the ecotype Col-0 also demonstrated for the first time the occurrence of AtGER3 protein in root extracts. Our results demonstrate the capacity of proteome analysis to analyse and distinguish closely related members of large protein families. PMID:15276453

  10. The expanding spectrum of COL2A1 gene variants IN 136 patients with a skeletal dysplasia phenotype.

    PubMed

    Barat-Houari, Mouna; Dumont, Bruno; Fabre, Aurélie; Them, Frédéric Tm; Alembik, Yves; Alessandri, Jean-Luc; Amiel, Jeanne; Audebert, Séverine; Baumann-Morel, Clarisse; Blanchet, Patricia; Bieth, Eric; Brechard, Marie; Busa, Tiffany; Calvas, Patrick; Capri, Yline; Cartault, François; Chassaing, Nicolas; Ciorca, Vidrica; Coubes, Christine; David, Albert; Delezoide, Anne-Lise; Dupin-Deguine, Delphine; El Chehadeh, Salima; Faivre, Laurence; Giuliano, Fabienne; Goldenberg, Alice; Isidor, Bertrand; Jacquemont, Marie-Line; Julia, Sophie; Kaplan, Josseline; Lacombe, Didier; Lebrun, Marine; Marlin, Sandrine; Martin-Coignard, Dominique; Martinovic, Jelena; Masurel, Alice; Melki, Judith; Mozelle-Nivoix, Monique; Nguyen, Karine; Odent, Sylvie; Philip, Nicole; Pinson, Lucile; Plessis, Ghislaine; Quélin, Chloé; Shaeffer, Elise; Sigaudy, Sabine; Thauvin, Christel; Till, Marianne; Touraine, Renaud; Vigneron, Jacqueline; Baujat, Geneviève; Cormier-Daire, Valérie; Le Merrer, Martine; Geneviève, David; Touitou, Isabelle

    2016-07-01

    Heterozygous COL2A1 variants cause a wide spectrum of skeletal dysplasia termed type II collagenopathies. We assessed the impact of this gene in our French series. A decision tree was applied to select 136 probands (71 Stickler cases, 21 Spondyloepiphyseal dysplasia congenita cases, 11 Kniest dysplasia cases, and 34 other dysplasia cases) before molecular diagnosis by Sanger sequencing. We identified 66 different variants among the 71 positive patients. Among those patients, 18 belonged to multiplex families and 53 were sporadic. Most variants (38/44, 86%) were located in the triple helical domain of the collagen chain and glycine substitutions were mainly observed in severe phenotypes, whereas arginine to cysteine changes were more often encountered in moderate phenotypes. This series of skeletal dysplasia is one of the largest reported so far, adding 44 novel variants (15%) to published data. We have confirmed that about half of our Stickler patients (46%) carried a COL2A1 variant, and that the molecular spectrum was different across the phenotypes. To further address the question of genotype-phenotype correlation, we plan to screen our patients for other candidate genes using a targeted next-generation sequencing approach. PMID:26626311

  11. Mutations in the COL5A1 gene are causal in the Ehlers-Danlos syndromes I and II

    SciTech Connect

    De Paepe, A.; Nuytinck, L.; Naeyaert, J.M.

    1997-03-01

    The Ehlers-Danlos syndrome (EDS) is a heterogeneous connective-tissue disorder of which at least nine subtypes are recognized. Considerable clinical overlap exists between the EDS I and II subtypes, suggesting that both are allelic disorders. Recent evidence based on linkage and transgenic mice studies suggest that collagen V is causally involved in human EDS. Collagen V forms heterotypic fibrils with collagen I in many tissues and plays an important role in collagen I fibrillogenesis. We have identified a mutation in COL5A1, the gene encoding the pro{alpha}1(V) collagen chain, segregating with EDS I in a four-generation family. The mutation causes the substitution of the most 5{prime} cysteine residue by a serine within a highly conserved sequence of the pro{alpha}1(V) C-propeptide domain and causes reduction of collagen V by preventing incorporation of the mutant pro{alpha}1 (V) chains in the collagen V trimers. In addition, we have detected splicing defects in the COL5A1 gene in a patient with EDS I and in a family with EDS II. These findings confirm the causal role of collagen V in at least a subgroup of EDS I, prove that EDS I and II are allelic conditions, and represent a, so far, unique example of a human collagen disorder caused by substitution of a highly conserved cysteine residue in the C-propeptide domain of a fibrillar collagen. 30 refs., 6 figs., 2 tabs.

  12. Carcinome à cellule vitreuse du col de l'utérus: à propos d'un cas et revue de littérature

    PubMed Central

    Hakimi, Ihssane; Zazi, Abdelghani; Chahdi, Hafsa; Guelzim, Khalid; Kouach, Jaouad; Babahabib, Myabdellah; Elhassani, Myehdi; Rahali, Driss Moussaoui; Dehayni, Mohammed

    2015-01-01

    Le carcinome à cellule vitreuse du col de l'utérus est un type de histologique rare de cancer du col de l'utérus qui survient à un âge plus jeune, et s'associe au risque élevé d’échec thérapeutique et le pronostic est plus mauvais en comparaison au type cellulaire squameux. La radiothérapie est associée au risque diminué de récidive. Le but de cette étude est de récapituler à travers d'une observation et une revue de littérature les données sur l'incidence, le comportement clinique et la survie globale de patients avec le carcinome à cellule vitreuse du col de l'utérus. PMID:26664556

  13. Complete structural organization of the human {alpha}1(V) collagen gene (COL5A1): Divergence from the conserved organization of other characterized fibrillar collagen genes

    SciTech Connect

    Takahara, Kazuhiko; Hoffman, G.G.; Greenspan, D.S.

    1995-10-10

    Genes that encode the vertebrate fibrillar collagen types I-III have previously been shown to share a highly conserved intron/exon organization, thought to reflect common ancestry and evolutionary pressures at the protein level. We report here the complete intron/exon organization of COL5A1, the human gene that encodes the {alpha}1 chain of fibrillar collagen type V. The structure of COL5A1 is shown to be considerably diverged from the conserved structure of the genes for fibrillar collagen types I-III. COL5A1 has 66 exons, which is greater than the number of exons found in the genes for collagen types I-III. The increased number of exons is partly due to the increased size of the pro-{alpha}1(V) N-propeptide, relative to the sizes of the N-propeptides of the types I-III procollagen molecules. In addition, however, the increased number of exons is due to differences in the intron/exon organization of the triple-helix coding region of COL5A1 compared to the organization of the triple-helix coding regions of the genes for collagen types I-III. Of particular interest is the increase of 54 bp exons in this region of COL5A1, strongly supporting the proposal that the triple-helix coding regions of fibrillar collagen genes evolved from duplication of a 54 bp primordial genetic element. Moreover, comparison of the structure of COL5A1 to the highly conserved structure of the genes of collagen types I-III provides insights into the probable structure of the ancestral gene that gave rise to what appears to be two classes of vertebrate fibrillar collagen genes. 50 refs., 5 figs.

  14. COL6A3 protein deficiency in mice leads to muscle and tendon defects similar to human collagen VI congenital muscular dystrophy.

    PubMed

    Pan, Te-Cheng; Zhang, Rui-Zhu; Markova, Dessislava; Arita, Machiko; Zhang, Yejia; Bogdanovich, Sasha; Khurana, Tejvir S; Bönnemann, Carsten G; Birk, David E; Chu, Mon-Li

    2013-05-17

    Collagen VI is a ubiquitously expressed extracellular microfibrillar protein. Its most common molecular form is composed of the α1(VI), α2(VI), and α3(VI) collagen α chains encoded by the COL6A1, COL6A2, and COL6A3 genes, respectively. Mutations in any of the three collagen VI genes cause congenital muscular dystrophy types Bethlem and Ullrich as well as intermediate phenotypes characterized by muscle weakness and connective tissue abnormalities. The α3(VI) collagen α chain has much larger N- and C-globular domains than the other two chains. Its most C-terminal domain can be cleaved off after assembly into microfibrils, and the cleavage product has been implicated in tumor angiogenesis and progression. Here we characterize a Col6a3 mutant mouse that expresses a very low level of a non-functional α3(VI) collagen chain. The mutant mice are deficient in extracellular collagen VI microfibrils and exhibit myopathic features, including decreased muscle mass and contractile force. Ultrastructurally abnormal collagen fibrils were observed in tendon, but not cornea, of the mutant mice, indicating a distinct tissue-specific effect of collagen VI on collagen I fibrillogenesis. Overall, the mice lacking normal α3(VI) collagen chains displayed mild musculoskeletal phenotypes similar to mice deficient in the α1(VI) collagen α chain, suggesting that the cleavage product of the α3(VI) collagen does not elicit essential functions in normal growth and development. The Col6a3 mouse mutant lacking functional α3(VI) collagen chains thus serves as an animal model for COL6A3-related muscular dystrophy. PMID:23564457

  15. cDNA cloning and chromosomal mapping of the mouse type VII collagen gene (Col7a1): Evidence for rapid evolutionary divergence of the gene

    SciTech Connect

    Li, Kehua; Christiano, A.M.; Chu, Mon Li; Uitto, J. Thomas Jefferson Univ., Philadelphia, PA ); Copeland, N.G.; Gilbert, D.J. )

    1993-06-01

    Type VII collagen is the major component of anchoring fibrils, critical attachment structures at the dermal-epidermal basement membrane zone. Genetic linkage analyses with recently cloned human type VII collagen cDNAs have indicated that the corresponding gene, COL7A1, is the candidate gene in the dystrophic forms of epidermolysis bullosa. To gain insight into the evolutionary conservation of COL7A1, in this study the authors have isolated mouse type VII collagen cDNAs by screening a mouse epidermal keratinocyte cDNA library with a human COL7A1 cDNA. Two overlapping mouse cDNAs were isolated, and Northern hybridization of mouse epidermal keratinocyte RNA with one of them revealed the presence of a mRNA transcript of [approximately]9.5 kb, the approximate size of the human COL7A1 mRNA. Nucleotide sequencing of the mouse cDNAs revealed a 2760-bp open reading frame that encodes the 5[prime] half of the collagenous domain and a segment of the NC-1, the noncollagenous amino-terminal domain of type VII collagen. Comparison of the mouse amino acid sequences with the corresponding human sequences deduced from cDNAs revealed 82.5% identity. The evolutionary divergence of the gene was relatively rapid in comparison to other collagen genes. Despite the high degree of sequence variation, several sequences, including the size and the position of noncollagenous imperfections and interruptions within the Gly-X-Y repeat sequence, were precisely conserved. Finally, the mouse Col7a1 gene was located by interspecific backcross mapping to mouse Chromosome 9, a region that corresponds to human chromosome 3p21, the position of human COL7Al. This assignment confirms and extends the relationship between the mouse and the human chromosomes in this region of the genome. 33 refs., 5 figs., 1 tab.

  16. Col6a1 Null Mice as a Model to Study Skin Phenotypes in Patients with Collagen VI Related Myopathies: Expression of Classical and Novel Collagen VI Variants during Wound Healing

    PubMed Central

    Lettmann, Sandra; Bloch, Wilhelm; Maaß, Tobias; Niehoff, Anja; Schulz, Jan-Niklas; Eckes, Beate; Eming, Sabine A.; Bonaldo, Paolo; Paulsson, Mats; Wagener, Raimund

    2014-01-01

    Patients suffering from collagen VI related myopathies caused by mutations in COL6A1, COL6A2 and COL6A3 often also display skin abnormalities, like formation of keloids or “cigarette paper” scars, dry skin, striae rubrae and keratosis pilaris (follicular keratosis). Here we evaluated if Col6a1 null mice, an established animal model for the muscle changes in collagen VI related myopathies, are also suitable for the study of mechanisms leading to the skin pathology. We performed a comprehensive study of the expression of all six collagen VI chains in unwounded and challenged skin of wild type and Col6a1 null mice. Expression of collagen VI chains is regulated in both skin wounds and bleomycin-induced fibrosis and the collagen VI α3 chain is proteolytically processed in both wild type and Col6a1 null mice. Interestingly, we detected a decreased tensile strength of the skin and an altered collagen fibril and basement membrane architecture in Col6a1 null mice, the latter being features that are also found in collagen VI myopathy patients. Although Col6a1 null mice do not display an overt wound healing defect, these mice are a relevant animal model to study the skin pathology in collagen VI related disease. PMID:25158062

  17. COL5A1: Fine genetic mapping, intron/exon organization, and exclusion as candidate gene in families with tuberous sclerosis complex 1, hereditary hemorrhagic telangiectasia, and Ehlers-Danlos syndrome type II

    SciTech Connect

    Greenspan, D.S.; Papenberg, K.A.; Marchuk, D.A.

    1994-09-01

    Type V collagen is the only fibrillar collagen which has yet to be implicated in the pathogenesis of genetic diseases in humans or mice. To begin examining the possible role of type V collagen in genetic disease, we have previously mapped COL5A1, the gene for the {alpha}1 chain of type V collagen, to 9q23.2{r_arrow}q34.3 and described two restriction site polymorphisms which allowed us to exclude COL5A1 as candidate gene for nail-patella syndrome. We have now used these polymorphisms to exclude COL5A1 as candidate gene for tuberous sclerosis complex 1 and Ehlers-Danlos syndrome type II. In addition, we describe a CA repeat, with observed heterozygosity of about 0.5, in a COL5A1 intron, which has allowed us to exclude COL5A1 as a candidate gene in hereditary hemorrhagic telangiectasia and to place COL5A1 on the CEPH family genetic map between markers D9S66 and D9S67. We have also determined the entire intron/exon organization of COL5A1, which will facilitate characterization of mutations in genetic diseases with which COL5A1 may be linked in future studies.

  18. Climate warming revealed by englacial temperatures at Col du Dôme (4250 m, Mont Blanc area)

    NASA Astrophysics Data System (ADS)

    Vincent, Christian; Le Meur, Emmanuel; Six, Delphine; Possenti, Philippe; Lefebvre, Eric; Funk, Martin

    2007-08-01

    Temperatures were measured in two deep boreholes drilled at the same location in the ice at Col du Dôme (4250 m) in 1994 and 2005, providing clear evidence of atmospheric warming. The 1994 temperature profile was already far from steady state conditions. Results from a heat transfer model reveal that the englacial temperature increase cannot be explained solely by atmospheric temperature rise. The latent heat produced by the refreezing of surface meltwater below the surface also contributes to the englacial temperature increase. Although surface melting is normally very low at this altitude, this contribution became significant after 1980 for temperatures at the top of the borehole. Simulations for different climatic scenarios show that glaciated areas located between 3500 and 4250 m could become temperate in the future. This warming could have a major impact on the stability of hanging glaciers frozen to their beds if the melting point is reached.

  19. Ehlers-Danlos syndrome, vascular type: a novel missense mutation in the COL3A1 gene.

    PubMed

    Masuno, Mitsuo; Watanabe, Atsushi; Naing, Banyar Than; Shimada, Takashi; Fujimoto, Wataru; Ninomiya, Shinsuke; Ueda, Yasunori; Kadota, Kazushige; Kotaka, Tatsuya; Kondo, Eisei; Yamanouchi, Yasuko; Inoue, Mika; Ouchi, Kazunobu; Kuroki, Yoshikazu

    2012-12-01

    We report a 34-year-old Japanese female with the vascular type of Ehlers-Danlos syndrome. She had thin translucent skin, extensive bruising, toe joint hypermobility, left lower extremity varicose veins, and chronic wrist, knee and ankle joint pain. She also had dizziness caused by autonomic dysfunction. Magnetic resonance angiography showed tortuous vertebral and basilar arteries, mild left carotid canal bulging, and right anterior tibial artery hypoplasia. Electron microscopic examinations of a skin biopsy revealed extremely dilated rough endoplasmic reticulum in dermal fibroblasts and wide variability of individual collagen fibril diameters. A molecular analysis using a conventional total RNA method and a high-resolution melting curve analysis using genomic DNA revealed a novel missense mutation within exon 48 of the COL3A1 gene, c.3428G>A, leading to p.Gly1143Glu. PMID:23181496

  20. Stickler-like syndrome due to a dominant negative mutation in the COL2A1 gene.

    PubMed

    Ballo, R; Beighton, P H; Ramesar, R S

    1998-10-30

    The type II collagenopathies include a wide spectrum of phenotypes ranging from mild spondylo epiphyseal dysplasia (SED) to severe achondrogenesis/hypochondrogenesis. Several attempts have been made at providing phenotype-genotype correlations in this group of disorders. In this report we discuss a South African family in which four members have a phenotype resembling Stickler syndrome type 1. Ocular problems and conductive deafness predominate, while skeletal changes resemble those of a mild form of multiple epiphyseal dysplasia (MED). In distinction to the classical form of Stickler syndrome, the affected persons have stubby digits. DNA analysis of the exons of the COL2A1 gene documented a C-T transversion in exon 39, resulting in an Arg704Cys substitution in the triple helical domain of the type II collagen peptide; this nontermination mutation may be indicative of further heterogeneity in the Stickler group of disorders or of a new syndrome amongst the type II collagenopathies. PMID:9800905

  1. [The Arabic influence in the "Colóquios dos simples e drogas da India" of Garcia da Orta].

    PubMed

    Ricordel, Joëlle

    2015-09-01

    The "Colóquios dos simples e drogas he cousas medicinais de Índia" (Conversations on the simples, drugs and medicinal substances of India) (1563) of Garcia da Orta is a botanical and pharmacognosy book. The author is a Portuguese physician who studied in the Spanish universities and practiced medicine mainly in India. He studies in short chapters presented in the form of dialogues about sixty simples. Sources to which he refers are indicative of a "classical" training, but also the mark of a curious and open mind to different cultures. The Arabic sources are numerous and mainly concern the identification of substances by abundant synonyms of their names in foreign languages and different medicinal uses that may have been done by the ancient physicians. However, Da Orta is critical with respect to these sources, seeking contradictions and differences of opinion among authors. He confronts them with the oral information collected thanks to a wide network of contacts. PMID:26529894

  2. Characterization of a type II collagen gene (COL2A1) mutation identified in cultured chondrocytes from human hypochondrogenesis.

    PubMed Central

    Horton, W A; Machado, M A; Ellard, J; Campbell, D; Bartley, J; Ramirez, F; Vitale, E; Lee, B

    1992-01-01

    A subtle mutation in the type II collagen gene COL2A1 was detected in a case of human hypochondrogenesis by using a chondrocyte culture system and PCR-cDNA scanning analysis. Chondrocytes obtained from cartilage biopsies were dedifferentiated and expanded in monolayer culture and then redifferentiated by culture over agarose. Single-strand conformation polymorphism and direct sequencing analysis identified a G----A transition, resulting in a glycine substitution at amino acid 574 of the pro alpha 1(II) collagen triple-helical domain. Morphologic assessment of cartilage-like structures produced in culture and electrophoretic analysis of collagens synthesized by the cultured chondrocytes suggested that the glycine substitution interferes with conversion of type II procollagen to collagen, impairs intracellular transport and secretion of the molecule, and disrupts collagen fibril assembly. This experimental approach has broad implications for the investigation of human chondrodysplasias as well as human chondrocyte biology. Images PMID:1374906

  3. SIN Retroviral Vectors Expressing COL7A1 Under Human Promoters for Ex Vivo Gene Therapy of Recessive Dystrophic Epidermolysis Bullosa

    PubMed Central

    Titeux, Matthias; Pendaries, Valérie; Zanta-Boussif, Maria A; Décha, Audrey; Pironon, Nathalie; Tonasso, Laure; Mejia, José E; Brice, Agnes; Danos, Olivier; Hovnanian, Alain

    2010-01-01

    Recessive dystrophic epidermolysis bullosa (RDEB) is caused by loss-of-function mutations in COL7A1 encoding type VII collagen which forms key structures (anchoring fibrils) for dermal–epidermal adherence. Patients suffer since birth from skin blistering, and develop severe local and systemic complications resulting in poor prognosis. We lack a specific treatment for RDEB, but ex vivo gene transfer to epidermal stem cells shows a therapeutic potential. To minimize the risk of oncogenic events, we have developed new minimal self-inactivating (SIN) retroviral vectors in which the COL7A1 complementary DNA (cDNA) is under the control of the human elongation factor 1α (EF1α) or COL7A1 promoters. We show efficient ex vivo genetic correction of primary RDEB keratinocytes and fibroblasts without antibiotic selection, and use either of these genetically corrected cells to generate human skin equivalents (SEs) which were grafted onto immunodeficient mice. We achieved long-term expression of recombinant type VII collagen with restored dermal–epidermal adherence and anchoring fibril formation, demonstrating in vivo functional correction. In few cases, rearranged proviruses were detected, which were probably generated during the retrotranscription process. Despite this observation which should be taken under consideration for clinical application, this preclinical study paves the way for a therapy based on grafting the most severely affected skin areas of patients with fully autologous SEs genetically corrected using a SIN COL7A1 retroviral vector. PMID:20485266

  4. Whole-Genome Sequence of Multidrug-Resistant Campylobacter coli Strain COL B1-266, Isolated from the Colombian Poultry Chain.

    PubMed

    Bernal, Johan F; Donado-Godoy, Pilar; Arévalo, Alejandra; Duarte, Carolina; Realpe, María E; Díaz, Paula L; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David; Mariño-Ramírez, Leonardo

    2016-01-01

    Campylobacter coli is considered one of the main causes of food-borne illness worldwide. We report here the whole-genome sequence of multidrug-resistant Campylobacter coli strain COL B1-266, isolated from the Colombian poultry chain. The genome sequences encode genes for a variety of antimicrobial resistance genes, including aminoglycosides, β-lactams, lincosamides, fluoroquinolones, and tetracyclines. PMID:26988047

  5. Whole-Genome Sequence of Multidrug-Resistant Campylobacter coli Strain COL B1-266, Isolated from the Colombian Poultry Chain

    PubMed Central

    Bernal, Johan F.; Donado-Godoy, Pilar; Arévalo, Alejandra; Duarte, Carolina; Realpe, María E.; Díaz, Paula L.; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David

    2016-01-01

    Campylobacter coli is considered one of the main causes of food-borne illness worldwide. We report here the whole-genome sequence of multidrug-resistant Campylobacter coli strain COL B1-266, isolated from the Colombian poultry chain. The genome sequences encode genes for a variety of antimicrobial resistance genes, including aminoglycosides, β-lactams, lincosamides, fluoroquinolones, and tetracyclines. PMID:26988047

  6. Cooperative interactions between PBX, PREP, and HOX proteins modulate the activity of the alpha 2(V) collagen (COL5A2) promoter.

    PubMed

    Penkov, D; Tanaka, S; Di Rocco, G; Berthelsen, J; Blasi, F; Ramirez, F

    2000-06-01

    Cell type-specific expression of the human alpha2(V) collagen (COL5A2) gene depends on a cis-acting element that consists of two contiguous protein binding sites (FPA and FPB) located between nucleotides -149 and -95, relative to the transcription start site. The present study focused on the characterization of the FPB-bound complex. DNA binding assays and cell transfection experiments revealed that the bipartite core sequence of FPB (5'-ATCAATCA-3') binds the PBX1/2, PREP1, and HOXB1 proteins, and this in turn leads to promoter transactivation. In the presence of all three nuclear factors, cooperative interactions between recombinant PBX1 and PREP1 or PBX1 and HOXB1 result in binding of the heterodimers to FPB in vitro. Similarly, overexpression of different combinations of PBX1, PREP1, and HOXB1 transactivates FPB-driven transcription. In contrast to the composition of the FPB complex purified from COL5A2-positive cells, the FPB complex from COL5A2-negative cells contains PBX2 and PREP1 but lacks PBX1. However, PBX1 exogenously introduced into COL5A2-negative cells cannot stimulate FPB-driven transcription unless co-expressed with PREP1. Within the intrinsic limitations of the experimental model, our results indicate that combinatorial interactions among PBX and PREP or HOX proteins are involved in regulating tissue-specific production of collagen V. PMID:10748126

  7. Identification of a novel COL2A1 mutation (c.1744G>A) in a Japanese family: a case report

    PubMed Central

    2014-01-01

    Introduction Mutations in the gene encoding the type II collagen gene (COL2A1) have been found to affect the entire skeletal system. Recently, inheritable skeletal dysplasia caused by novel COL2A1 mutations has been linked to an inherited disease of the hip joint that neither involves the entire skeletal system nor is characterized by the presence of concomitant disorders, such as spinal or ocular abnormalities. Case presentation A 27-year-old Japanese woman previously diagnosed with avasucular necrosis (AVN) of the femoral head on the basis of radiological findings was referred to the study site for surgical management of a painful hip joint. She had no history of disease but suffered from bilateral hip joint lesions. Analysis of her pedigree revealed that bilateral hip joint lesions affected more than three generations of her family. Based on these findings, haplotype analysis of her and her family members was performed by examining select candidate genes from the critical interval for epiphyseal dysplasia of the femoral head on 12q13 and sequencing the promoter and exonic regions of COL2A1. Conclusion A novel COL2A1 mutation (c.1744G>A) was identified within one Japanese family. PMID:25124518

  8. Novel X-linked glomerulopathy is associated with a COL4A5 missense mutation in a non-collagenous interruption.

    PubMed

    Becknell, Brian; Zender, Gloria A; Houston, Ronald; Baker, Peter B; McBride, Kim L; Luo, Wentian; Hains, David S; Borza, Dorin-Bogdan; Schwaderer, Andrew L

    2011-01-01

    A novel COL4A5 mutation causes rapid progression to end-stage renal disease in males, despite the absence of clinical and biopsy findings associated with Alport syndrome. Affected males have proteinuria, variable hematuria, and an early progression to end-stage renal disease. Renal biopsy findings include global and segmental glomerulosclerosis, mesangial hypercellularity and basement membrane immune complex deposition. Exon sequencing of the COL4A5 locus identified a thymine to guanine transversion at nucleotide 665, resulting in a phenylalanine to cysteine missense mutation at codon 222. The phenylalanine at position 222 is absolutely conserved among vertebrates. This mutation was confirmed in 4 affected males and 4 female obligate carriers, but was absent in 6 asymptomatic male family members and 198 unrelated individuals. Immunostaining for α5(IV) collagen in renal biopsies from affected males was normal. This mutation, in a non-collagenous interruption associated with severe renal disease, provides evidence for the importance of this structural motif and suggests the range of phenotypes associated with COL4A5 mutations is more diverse than previously realized. Hence, COL4A5 mutation analysis should be considered when glomerulonephritis presents in an X-linked inheritance pattern, even with a presentation distinct from Alport syndrome. PMID:20881942

  9. Distribution of ColE1-like KAN-resistance plasmids in a population of Salmonella enterica from animal diagnostic samples

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Previous studies showed that some Salmonella isolates may harbor ColE1-like plasmids that carry a single aph gene responsible for kanamycin resistance (KAN-R). However the distribution of these plasmids is unknown. Methods: KAN-R Salmonella isolates (n=102) collected through the 2005 N...

  10. Genetic linkage of type VII collagen (COL7A1) to dominant dystrophic epidermolysis bullosa in families with abnormal anchoring fibrils.

    PubMed Central

    Ryynänen, M; Ryynänen, J; Sollberg, S; Iozzo, R V; Knowlton, R G; Uitto, J

    1992-01-01

    Epidermolysis bullosa (EB) in a group of genodermatoses characterized by the fragility of skin. Previous studies on the dystrophic (scarring) forms of EB have suggested abnormalities in anchoring fibrils, morphologically recognizable attachment structures that provide stability to the association of the cutaneous basement membrane to the underlying dermis. Since type VII collagen is the major component of the anchoring fibrils, we examined the genetic linkage of dominant dystrophic EB (EBDD) and the type VII collagen gene (COL7A1) locus, which we have recently mapped to chromosome 3p, in three large kindreds with abnormal anchoring fibrils. Strong genetic linkage of EBDD and COL7A1 loci was demonstrated with the maximum logarithm of odds (LOD) score of 8.77 at theta = 0. This linkage was further confirmed with two additional markers in this region of the short arm of chromosome 3, and these analyses allowed further refinement of the map locus of COL7A1. Since there were no recombinants between the COL7A1 and EBDD loci, our findings suggest that type VII collagen is the candidate gene that may harbor the mutations responsible for the EB phenotype in these three families. Images PMID:1347297

  11. The Tsk2/+ mouse fibrotic phenotype is due to a gain-of-function mutation in the PIIINP segment of the Col3a1 gene.

    PubMed

    Long, Kristen B; Li, Zhenghui; Burgwin, Chelsea M; Choe, Susanna G; Martyanov, Viktor; Sassi-Gaha, Sihem; Earl, Josh P; Eutsey, Rory A; Ahmed, Azad; Ehrlich, Garth D; Artlett, Carol M; Whitfield, Michael L; Blankenhorn, Elizabeth P

    2015-03-01

    Systemic sclerosis (SSc) is a polygenic, autoimmune disorder of unknown etiology, characterized by the excessive accumulation of extracellular matrix (ECM) proteins, vascular alterations, and autoantibodies. The tight skin (Tsk)2/+ mouse model of SSc demonstrates signs similar to SSc including tight skin and excessive deposition of dermal ECM proteins. By linkage analysis, we mapped the Tsk2 gene mutation to <3 megabases on chromosome 1. We performed both RNA sequencing of skin transcripts and genome capture DNA sequencing of the region spanning this interval in Tsk2/+ and wild-type littermates. A missense point mutation in the procollagen III amino terminal propeptide segment (PIIINP) of collagen, type III, alpha 1 (Col3a1) was found to be the best candidate for Tsk2; hence, both in vivo and in vitro genetic complementation tests were used to prove that this Col3a1 mutation is the Tsk2 gene. All previously documented mutations in the human Col3a1 gene are associated with the Ehlers-Danlos syndrome, a connective tissue disorder that leads to a defect in type III collagen synthesis. To our knowledge, the Tsk2 point mutation is the first documented gain-of-function mutation associated with Col3a1, which leads instead to fibrosis. This discovery provides insight into the mechanism of skin fibrosis manifested by Tsk2/+ mice. PMID:25330296

  12. Characterization of mutations of the type VII collagen gene (COL7A1) in recessive dystrophic epidermolysis bullosa mitis (M-RDEB) from three Korean patients.

    PubMed

    Ryoo, Y W; Kim, B C; Lee, K S

    2001-06-01

    In recent years, the molecular basis for the main subtypes of epidermolysis bullosa (EB) has been elucidated with pathogenetic mutations delineated in ten different genes encoding structural components of the dermal-epidermal junction. Both the autosomal dominant and recessive forms of dystrophic EB (DEB) is caused by mutations in the COL7A1 gene. Type VII collagen is a major component of anchoring fibrils, structural elements that stabilize the attachment of the basement membrane to underlying dermis. Recent delineation of the exon-intron organization of the COL7A1 gene provided the basis for the comprehensive design of PCR primer pairs that amplified exons in genomic DNA by placing the primers on the flanking introns. A number of COL7A1 mutations have been reported and some genotype-phenotype correlations are starting to emerge. In this study, we examined mutational analyses from three Korean patients with recessive dystrophic EB (RDEB) mitis. We designed and optimized primers according to the previously reported sequences. Such PCR amplification products can be examined by electrophoretic scanning technique, CSGE heteroduplex analyses. Utilizing heteroduplex analyses, we have identified a number of sequence variants in COL7A1 both in unaffected individuals and in patients with M-RDEB. Mutation detection of the COL7A1 gene revealed six allelic mutations (V6677E, P6685S, Y3749S, P6084S, P6695R and G6697C). We suggest that the full length of type VII collagen polypeptide are synthesized, but those missense mutations, that may affect a critical amino acid, can alter the conformation of the protein and interferes with the assembly and packing of type VII collagen molecules into anchoring fibrils. Immunohistochemical study of skin biopsies by use of anti-type VII collagen antibody showed markedly reduced staining and presence of a dermo/epidermal cleavage. This is the first report of a COL7A1 mutation study in DEB from Korean patients. We hope that these data

  13. A Novel Splice Variant in the N-propeptide of COL5A1 Causes an EDS Phenotype with Severe Kyphoscoliosis and Eye Involvement

    PubMed Central

    Symoens, Sofie; Malfait, Fransiska; Vlummens, Philip; Hermanns-Lê, Trinh; Syx, Delfien; De Paepe, Anne

    2011-01-01

    Background The Ehlers-Danlos Syndrome (EDS) is a heritable connective tissue disorder characterized by hyperextensible skin, joint hypermobility and soft tissue fragility. The classic subtype of EDS is caused by mutations in one of the type V collagen genes (COL5A1 and COL5A2). Most mutations affect the type V collagen helical domain and lead to a diminished or structurally abnormal type V collagen protein. Remarkably, only two mutations were reported to affect the extended, highly conserved N-propeptide domain, which plays an important role in the regulation of the heterotypic collagen fibril diameter. We identified a novel COL5A1 N-propeptide mutation, resulting in an unusual but severe classic EDS phenotype and a remarkable splicing outcome. Methodology/Principal Findings We identified a novel COL5A1 N-propeptide acceptor-splice site mutation (IVS6-2A>G, NM_000093.3_c.925-2A>G) in a patient with cutaneous features of EDS, severe progressive scoliosis and eye involvement. Two mutant transcripts were identified, one with an exon 7 skip and one in which exon 7 and the upstream exon 6 are deleted. Both transcripts are expressed and secreted into the extracellular matrix, where they can participate in and perturb collagen fibrillogenesis, as illustrated by the presence of dermal collagen cauliflowers. Determination of the order of intron removal and computational analysis showed that simultaneous skipping of exons 6 and 7 is due to the combined effect of delayed splicing of intron 7, altered pre-mRNA secondary structure, low splice site strength and possibly disturbed binding of splicing factors. Conclusions/Significance We report a novel COL5A1 N-propeptide acceptor-splice site mutation in intron 6, which not only affects splicing of the adjacent exon 7, but also causes a splicing error of the upstream exon 6. Our findings add further insights into the COL5A1 splicing order and show for the first time that a single COL5A1 acceptor-splice site mutation can perturb

  14. The Variants Within the COL5A1 Gene are Associated with Reduced Risk of Anterior Cruciate Ligament Injury in Skiers.

    PubMed

    Stępień-Słodkowska, Marta; Ficek, Krzysztof; Kaczmarczyk, Mariusz; Maciejewska-Karłowska, Agnieszka; Sawczuk, Marek; Leońska-Duniec, Agata; Stępiński, Miłosz; Ziętek, Paweł; Król, Paweł; Chudecka, Monika; Cięszczyk, Paweł

    2015-03-29

    The purpose of this study was to examine the association of the BstUI RFLP C/T (rs 12722) and DpnII RFLP C/T (rs 13946) COL5A1 polymorphisms, individually and as haplotypes, with anterior cruciate ligament ruptures in recreational skiers. Subjects were 138 male recreational skiers with surgically diagnosed primary anterior cruciate ligament ruptures. The control group consisted of 183 apparently healthy male recreational skiers, who were without any self-reported history of ligament or tendon injury. DNA was extracted from buccal cells donated by the subjects and genotyping was carried out using real-time PCR. The genotype distributions for both polymorphisms met Hardy-Weinberg expectations in both groups. There were no significant differences in genotype distribution of allele frequencies of COL5A1 BstUI RFLP C/T and COL5A1 DpnII RFLP C/T polymorphisms between the ACL rupture and control groups. The T-T (BstUI RFLP T, DpnII RFLP T) haplotype was the most common (55.6%). The haplotype T-C was not present in any of the subjects. There was an underrepresentation tendency of the C-T haplotype in the study group compared to controls under recessive mode of inheritance. Higher frequency of the COL5A1 BstUI RFLP C/T and COL5A1DpnII RFLP C/T polymorphisms haplotype is associated with reduced risk of anterior cruciate ligament injury in a group of apparently healthy male recreational skiers. PMID:25964814

  15. Characterization of 18 new mutations in COL7A1 in recessive dystrophic epidermolysis bullosa provides evidence for distinct molecular mechanisms underlying defective anchoring fibril formation.

    PubMed Central

    Hovnanian, A; Rochat, A; Bodemer, C; Petit, E; Rivers, C A; Prost, C; Fraitag, S; Christiano, A M; Uitto, J; Lathrop, M; Barrandon, Y; de Prost, Y

    1997-01-01

    We have characterized 21 mutations in the type VII collagen gene (COL7A1) encoding the anchoring fibrils, 18 of which were not previously reported, in patients from 15 unrelated families with recessive dystrophic epidermolysis bullosa (RDEB). COL7A1 mutations in both alleles were identified by screening the 118 exons of COL7A1 and flanking intron regions. Fourteen mutations created premature termination codons (PTCs) and consisted of nonsense mutations, small insertions, deletions, and splice-site mutations. A further seven mutations predicted glycine or arginine substitutions in the collagenous domain of the molecule. Two mutations were found in more than one family reported in this study, and six of the seven missense mutations showed clustering within exons 72-74 next to the hinge region of the protein. Patients who were homozygous or compound heterozygotes for mutations leading to PTCs displayed both absence or drastic reduction of COL7A1 transcripts and undetectable type VII collagen protein in skin. In contrast, missense mutations were associated with clearly detectable COL7A1 transcripts and with normal or reduced expression of type VII collagen protein at the dermo/epidermal junction. Our results provide evidence for at least two distinct molecular mechanisms underlying defective anchoring fibril formation in RDEB: one involving PTCs leading to mRNA instability and absence of protein synthesis, the other implicating missense mutations resulting in the synthesis of type VII collagen polypeptide with decreased stability and/or altered function. Genotype-phenotype correlations suggested that the nature and location of these mutations are important determinants of the disease phenotype and showed evidence for interfamilial phenotypic variability. Images Figure 1 Figure 2 Figure 4 Figure 5 Figure 6 PMID:9326325

  16. Clinical, structural, biochemical and X-ray crystallographic correlates of pathogenicity for variants in the C-propeptide region of the COL3A1 gene.

    PubMed

    Stembridge, Natasha S; Vandersteen, Anthony M; Ghali, Neeti; Sawle, Philip; Nesbitt, Mandy; Pollitt, Rebecca C; Ferguson, David J P; Holden, Simon; Elmslie, Frances; Henderson, Alex; Hulmes, David J S; Pope, F Michael

    2015-08-01

    Vascular Ehlers-Danlos syndrome (vEDS) is a heritable disorder of connective tissue caused by pathological variants in the COL3A1 gene, which encodes the α1 chain of type III collagen. Type III collagen is a major component of skin, arterial walls, and the gastrointestinal tract. Collagen III protein deficiency manifests as an increased risk of rupture, perforation, and dissection of these structures. The most disruptive gene variants affect the collagen helix via glycine substitutions or splice donor site mutations. The C-propeptide region of COL3A1 includes exons 49-52 and has a crucial role in initiating the C-terminal assembly of procollagen monomers in the early stages of collagen biosynthesis. Nineteen COL3A1 variants have previously been reported in these exons, of which four were associated with a severe vEDS phenotype. We identified two novel C-propeptide missense variants; p.Pro1440Leu, p.Arg1432Leu, and a non-stop mutation, c.4400A > T, p. (*1467Leuext*45). These variants produce variable phenotypes ranging from obvious acrogeria to classical or hypermobile EDS. A previously reported variant p.Lys1313Arg is of unknown clinical significance but likely benign, based on this study. Assigning disease pathogenicity remains complex, clinical phenotyping and crystal structure evidence being crucial. We briefly compare reported phenotypes for patients with missense variants in the C-propeptide domain for other human collagen disorders including COL1A1 and COL1A2 (osteogenesis imperfecta). PMID:25846194

  17. DEVELOPMENTAL AND OSTEOARTHRITIC CHANGES IN Col6a1 KNOCKOUT MICE: THE BIOMECHANICS OF COLLAGEN VI IN THE CARTILAGE PERICELLULAR MATRIX

    PubMed Central

    Alexopoulos, Leonidas G.; Youn, Inchan; Bonaldo, Paolo; Guilak, Farshid

    2009-01-01

    Chondrocytes are the sole cell type in articular cartilage and maintain the extracellular matrix through a homeostatic balance of anabolic and catabolic activities that are influenced by genetic factors, soluble mediators, and biophysical factors such as mechanical stress. Chondrocytes are encapsulated by a narrow tissue region termed the “pericellular matrix”, which in normal cartilage is defined by the exclusive presence of type VI collagen. Because the pericellular matrix completely surrounds each cell, it is hypothesized to serve as a filter or transducer for biochemical and/or biomechanical signals from the cartilage extracellular matrix. In this study, we used Col6a1−/− mice to investigate whether the lack of collagen VI may affect the development and biomechanical function of the pericellular matrix and alter the mechanical environment of the chondrocytes during joint loading. Col6a1−/− and Col6a1+/− mice possessed structurally intact pericellular matrices, but with significantly reduced mechanical properties as compared to wild-type controls. With age, Col6a1−/− showed accelerated development of osteoarthritic joint degeneration, as well as other musculoskeletal abnormalities such as delayed secondary ossification process and reduced bone mineral density. These findings suggest an important role for type VI collagen in regulating the physiology of the synovial joint, and provide indirect evidence that alterations in the mechanical environment of the chondrocytes, either due to loss of pericellular matrix properties or Col6α1−/− derived joint laxity, can lead to the progression of osteoarthritis. PMID:19248115

  18. Analysis of four families with the Stickler syndrome by linkage studies. Identification of a new premature stop codon in the COL2A1 gene in a family

    SciTech Connect

    Bonaventure, J.; Lasselin, C.; Toutain, A.

    1994-09-01

    The Stickler syndrome is an arthro-ophthalmopathy which associates progressive myopia with vitreal degeneration and retinal detachment. Cleft palate, cranio-facial abnormalities, deafness and osteoarthritis are often associated symptoms. Genetic heterogeneity of this autosomal dominant disease was consistent with its large clinical variability. Linkage studies have provided evidence for cosegregation of the disease with COL2A1, the gene coding for type II collagen, in about 50% of the families. Four additional families are reported here. Linkage analyses by using a VNTR located in the 3{prime} region of the gene were achieved. In three families, positive lod scores were obtained with a cumulative maximal value of 3.5 at a recombination fraction of 0. In one of these families, single strand conformation analysis of 25 exons disclosed a new mutation in exon 42. Codon for glutamic acid at position a1-803 was converted into a stop codon. The mutation was detected in DNA samples from all the affected members of the family but not in the unaffected. This result confirms that most of the Stickler syndromes linked to COL2A1 are due to premature stop codons. In a second family, an abnormal SSCP pattern of exon 34 was detected in all the affected individuals. The mutation is likely to correspond to a splicing defect in the acceptor site of intron 33. In one family the disease did not segregate with the COL2A1 locus. Further linkage studies with intragenic dimorphic sites in the COL10A1 gene and highly polymorphic markers close to the COL9A1 locus indicated that this disorder did not result from defects in these two genes.

  19. COL5A1: Genetic mapping and exclusion as candidate gene in families with nail-patella syndrome, tuberous sclerosis 1, hereditary hemorrhagic telangiectasia, and Ehlers-Danlos syndrome type II

    SciTech Connect

    Greenspan, D.S.; Northrup, H.; Au, K.S.

    1995-02-10

    COL5A1, the gene for the {alpha}1 chain of type V collagen, has been considered a candidate gene for certain diseases based on chromosomal location and/or disease phenotype. We have employed 3{prime}-untranslated region RFLPs to exclude COL5A1 as a candidate gene in families with tuberous sclerosis 1, Ehlers-Danlos syndrome type H, and nail-patella syndrome. In addition, we describe a polymorphic simple sequence repeat (SSR) within a COL5A1 intron. This SSR is used to exclude COL5A1 as a candidate gene in hereditary hemorrhagic telangiectasia (Osler-Rendu-Weber disease) and to add COL5A1 to the existing map of {open_quotes}index{close_quotes} markers of chromosome 9 by evaluation of the COL5A1 locus on the CEPH 40-family reference pedigree set. This genetic mapping places COL5A1 between markers D9S66 and D9S67. 14 refs., 1 fig., 2 tabs.

  20. A CONSTANS-like transcriptional activator, OsCOL13, functions as a negative regulator of flowering downstream of OsphyB and upstream of Ehd1 in rice.

    PubMed

    Sheng, Peike; Wu, Fuqing; Tan, Junjie; Zhang, Huan; Ma, Weiwei; Chen, Liping; Wang, Jiachang; Wang, Jie; Zhu, Shanshan; Guo, Xiuping; Wang, Jiulin; Zhang, Xin; Cheng, Zhijun; Bao, Yiqun; Wu, Chuanyin; Liu, Xuanming; Wan, Jianmin

    2016-09-01

    Flowering time determines the adaptability of crop plants to different local environments, thus being one of the most important agronomic traits targeted in breeding programs. Photoperiod is one of the key factors that control flowering in plant. A number of genes that participate in the photoperiod pathway have been characterized in long-day plants such as Arabidopsis, as well as in short-day plants such as Oryza sativa. Of those, CONSTANS (CO) as a floral integrator promotes flowering in Arabidopsis under long day conditions. In rice, Heading date1 (Hd1), a homologue of CO, functions in an opposite way, which inhibits flowering under long day conditions and induces flowering under short day conditions. Here, we show that another CONSTANS-like (COL) gene, OsCOL13, negatively regulates flowering in rice under both long and short day conditions. Overexpression of OsCOL13 delays flowering regardless of day length. We also demonstrated that OsCOL13 has a constitutive and rhythmic expression pattern, and that OsCOL13 is localized to the nucleus. OsCOL13 displays transcriptional activation activity in the yeast assays and likely forms homodimers in vivo. OsCOL13 suppresses the florigen genes Hd3a and RFT1 by repressing Ehd1, but has no relationship with other known Ehd1 regulators as determined by using mutants or near isogenic lines. In addition, the transcriptional level of OsCOL13 significantly decreased in the osphyb mutant, but remained unchanged in the osphya and osphyc mutants. Thus, we conclude that OsCOL13 functions as a negative regulator downstream of OsphyB and upstream of Ehd1 in the photoperiodic flowering in rice. PMID:27405463

  1. Environmental conditions at the South Col of Mount Everest and their impact on hypoxia and hypothermia experienced by mountaineers

    PubMed Central

    2012-01-01

    Background Hypoxia and hypothermia are acknowledged risk factors for those who venture into high-altitude regions. There is, however, little in situ data that can be used to quantify these risks. Here, we use 7 months of continuous meteorological data collected at the South Col of Mount Everest (elevation 7,896 m above sea level) to provide the first in situ characterization of these risks near the summit of Mount Everest. Methods This is accomplished through the analysis of barometric pressure, temperature and wind speed data collected by an automatic weather station installed at the South Col. These data were also used as inputs to parameterizations of wind chill equivalent temperature (WCT) and facial frostbite time (FFT). Results The meteorological data show clear evidence of seasonality, with evidence of pre-monsoon, monsoon and post-monsoon conditions. Low pressures, cold temperatures and high wind speeds characterize the pre- and post-monsoon periods with significant variability associated with the passage of large-scale weather systems. In contrast, the monsoon period is characterized by higher pressures, warmer temperatures and lower wind speeds with a pronounced reduction in variability. These environmental conditions are reflected in WCTs as low as −50°C and FFTs as short as 2 min during the pre- and post-monsoon periods. During the monsoon, the risk of cold injury is reduced with WCTs of order −20°C and FFTs longer than 60 min. The daily cycle in the various parameters is also investigated in order to assess the changes in conditions that would be experienced during a typical summit day. The post-monsoon period in particular shows a muted daily cycle in most parameters that is proposed to be the result of the random timing of large-scale weather systems. Conclusions Our results provide the first in situ characterization of the risk of hypoxia and hypothermia on Mount Everest on daily, weekly and seasonal timescales, and provide additional

  2. Tailoring the interfacial exchange coupling of perpendicularly magnetized Co/L10-Mn1.5Ga bilayers

    NASA Astrophysics Data System (ADS)

    Xiao, J. X.; Lu, J.; Liu, W. Q.; Zhang, Y. W.; Wang, H. L.; Zhu, L. J.; Deng, H. X.; Wei, D. H.; Xu, Y. B.; Zhao, J. H.

    2016-06-01

    We have studied the magnetic properties of Co (2–12 MLs)/L10-Mn1.5Ga (15 nm) bilayers without and with annealing at 300 °C by a combination of superconducting quantum interference device (SQUID) magnetometry and x-ray magnetic circular dichroism (XMCD). We find that the Co layer can remain perpendicularly magnetized when its thickness is less than six monolayers due to the coupling between Co and L10-Mn1.5Ga layers, which is doubly confirmed by both SQUID and XMCD measurements. Such an exchange coupling between L10-Mn1.5Ga and Co layers changes from ferromagnetic coupling to antiferromagnetic coupling after the annealing process. Furthermore, the magnetic coupling can also be tailored from ferromagnetic to antiferromagnetic by changing the L10-Mn1.5Ga surface from Mn-rich to Ga-rich. The first-principles calculations show that the interfacial coupling type is ferromagnetic for a Mn-terminated L10-Mn1.5Ga bilayer, while antiferromagnetic for a Ga-terminated bilayer. The spin and orbital moments of Co in the Co/L10-Mn1.5Ga bilayers are calculated according to the sum rules and the ratio of the orbital to spin magnetic moments for Co is not enhanced like other perpendicularly magnetized Co-based multilayers such as Co/Pd and Co/Pt.

  3. Variable clinical manifestations of a glycine to glutamic acid substitution of the COL3A1 gene at residue 736

    SciTech Connect

    Pope, F.M.; Narcisi, P.; Richards, A.J.

    1994-09-01

    Glycine substitutions at the 3{prime} end of the COL3A1 gene generally produce a characteristic clinical phenotype including acrogeria and severe vascular fragility. Here we report a three generation British family in which the propositus presented with aneurysms of the groins. He, his mother, sister and elder daughter all had the external clinical phenotype of vascular EDS IV whilst another daughter and nephew were clinically normal. Cultured skin fibroblasts from the propositus and his clinically affected relatives poorly secreted normal and overmodified collagen III species. Normal components of secreted proteins predominated whilst overmodified molecules were prominent in intracellular material. Surprisingly the normal children also secreted less collagen type III than expected (though more than their clinically abnormal relatives). cDNA from bases 2671 to 3714 were amplified as four overlapping PCR fragments and analysed by DGGE. The region between 2671 and 3015 was heterozygous. Sequencing showed a mutation of glycine to glutamic acid at residue 736. This mutation created an extra Apa 1 restriction site which was suitable for family studies. These showed inheritance of the mutant gene by both vascular and non-vascular clinical phenotypes. This family therefore illustrates that replacement of glycine to glutamic acid at position 736 produces variable clinical and biochemical phenotypes ranging from easily recognizable vascular EDS IV with very poor collagen secretion to an EDS III-like picture and with less severe protein disturbance. The reasons for these differences are at present unexplained.

  4. The first Japanese case of the arthrochalasia type of Ehlers-Danlos syndrome with COL1A2 gene mutation.

    PubMed

    Hatamochi, Atsushi; Hamada, Takahiro; Yoshino, Makoto; Hashimoto, Takashi

    2014-03-15

    This is the first report for a Japanese case of arthrochalasia type of Ehlers-Danlos syndrome (EDS). A 46-year-old woman consulted us for joint hypermobility and skin hyperextensibility that had been present soon after birth. There was no family history of a similar disease. She was diagnosed as having bilateral congenital hip dislocation and bilateral habitual shoulder dislocation at her childhood. Her skin was velvety, doughy and hyperextensible. She showed hypermobility of the joints of the hands and feet and generalized joint laxity, with no evidence of scoliosis. Electrophoretic analysis of collagenous proteins revealed the presence of an additional band in the position of pNα2(I) in the sample from culture medium of the patient fibroblasts. Analysis of the α2 chains of type I collagen gene, COL1A2, showed a heterozygous G to T transition at the +1 position of the exon 6 donor splice site (c.279+1G>T). This mutation resulted in skipping of exon 6, which leads to deficient processing of the amino-terminal end of proα2(I) chains of type I collagen. Based on these findings, we made a diagnosis of the arthrochalasia type of EDS, which corresponds to EDS type VIIB in the former classification. PMID:24440294

  5. COL11A2 mutation associated with autosomal recessive Weissenbacher-Zweymuller syndrome: molecular and clinical overlap with otospondylomegaepiphyseal dysplasia (OSMED).

    PubMed

    Harel, Tamar; Rabinowitz, Ronen; Hendler, Netta; Galil, Aharon; Flusser, Hagit; Chemke, Juan; Gradstein, Libe; Lifshitz, Tova; Ofir, Rivka; Elbedour, Khalil; Birk, Ohad S

    2005-01-01

    Autosomal recessive Weissenbacher-Zweymuller syndrome (WZS) is a skeletal dysplasia characterized by rhizomelic dwarfism and severe hearing loss. Mutations in the COL11A2 gene have been implicated in causing the autosomal dominant form of this syndrome as well as non-ocular Stickler syndrome and the autosomal recessive syndrome otospondylomegaepiphyseal dysplasia (OSMED). In a consanguineous Bedouin tribe living in Southern Israel, five individuals affected by autosomal recessive WZS were available for genetic analysis. Homozygosity of a mutation in the COL11A2 gene was found in all affected individuals. This finding lends molecular support to the clinical notion that autosomal recessive WZS and OSMED are a single entity. PMID:15558753

  6. Co-Occurence of Reciprocal Translocation and COL2A1 Mutation in a Fetus with Severe Skeletal Dysplasia: Implications for Genetic Counseling.

    PubMed

    Heinrich, Tilman; Nanda, Indrajit; Rehn, Monika; Zollner, Ursula; Ernestus, Karen; Wirth, Clemens; Schlüter, Gregor; Schmid, Michael; Kunstmann, Erdmute

    2015-01-01

    Achondrogenesis type II is an autosomal-dominant disease leading to severe micromelic dwarfism. Here, we report on the postmortem identification of a de novo heterozygous mutation in the COL2A1 gene (c.1529G>A, p.Gly510Asp) in a fetus who presented with generalized hydrops fetalis and severe micromelia during prenatal sonographic examinations. Initially, a reciprocal translocation t(4;17)(q31;p13) was detected in this fetus by chorionic villus sampling. Subsequent chromosomal analysis of maternal and paternal blood showed that the patient's mother was carrier of the same reciprocal translocation. SNP array analysis of the fetus did not provide evidence for chromosomal imbalances or CNVs that could be associated with the fetal phenotype. The coexistence of a cytogenetic (reciprocal translocation) and a molecular genetic (COL2A1 mutation) abnormality in the fetus carries important implications for genetic counseling. PMID:25823796

  7. Transformation of Shewanella baltica with ColE1-like and P1 plasmids and their maintenance during bacterial growth in cultures.

    PubMed

    Milewska, Klaudia; Węgrzyn, Grzegorz; Szalewska-Pałasz, Agnieszka

    2015-09-01

    The presence of natural plasmids has been reported for many Shewanella isolates. However, knowledge about plasmid replication origin and segregation mechanisms is not extensive for this genus. Shewanella baltica is an important species in the marine environment due to its denitrification ability in oxygen-deficient zones and the potential role in bioremediation processes. However, no information about possible use of plasmid vectors in this species has been reported to date. Here we report that plasmids with ColE1-type and plasmid P1 origin can transform S. baltica and replicate in this bacterium. Without the antibiotic selection pressure plasmid maintenance is less efficient than in Escherichia coli. Nevertheless, cultivation of S. baltica in the presence of appropriate antibiotics caused relatively stable maintenance of ColE1-like and P1-derived plasmids. This indicates that plasmid-based genetic manipulations and gene transfer in S. baltica are possible. PMID:26170108

  8. Identification of a TAAT-containing motif required for high level expression of the COL1A1 promoter in differentiated osteoblasts of transgenic mice

    NASA Technical Reports Server (NTRS)

    Dodig, M.; Kronenberg, M. S.; Bedalov, A.; Kream, B. E.; Gronowicz, G.; Clark, S. H.; Mack, K.; Liu, Y. H.; Maxon, R.; Pan, Z. Z.; Upholt, W. B.; Rowe, D. W.; Lichtler, A. C.

    1996-01-01

    Our previous studies have shown that the 49-base pair region of promoter DNA between -1719 and -1670 base pairs is necessary for transcription of the rat COL1A1 gene in transgenic mouse calvariae. In this study, we further define this element to the 13-base pair region between -1683 and -1670. This element contains a TAAT motif that binds homeodomain-containing proteins. Site-directed mutagenesis of this element in the context of a COL1A1-chloramphenicol acetyltransferase construct extending to -3518 base pairs decreased the ratio of reporter gene activity in calvariae to tendon from 3:1 to 1:1, suggesting a preferential effect on activity in calvariae. Moreover, chloramphenicol acetyltransferase-specific immunofluorescence microscopy of transgenic calvariae showed that the mutation preferentially reduced levels of chloramphenicol acetyltransferase protein in differentiated osteoblasts. Gel mobility shift assays demonstrate that differentiated osteoblasts contain a nuclear factor that binds to this site. This binding activity is not present in undifferentiated osteoblasts. We show that Msx2, a homeodomain protein, binds to this motif; however, Northern blot analysis revealed that Msx2 mRNA is present in undifferentiated bone cells but not in fully differentiated osteoblasts. In addition, cotransfection studies in ROS 17/2.8 osteosarcoma cells using an Msx2 expression vector showed that Msx2 inhibits a COL1A1 promoter-chloramphenicol acetyltransferase construct. Our results suggest that high COL1A1 expression in bone is mediated by a protein that is induced during osteoblast differentiation. This protein may contain a homeodomain; however, it is distinct from homeodomain proteins reported previously to be present in bone.

  9. Association analysis of the COL1A1 polymorphism with bone mineral density and prevalent fractures in Polish postmenopausal women with osteoporosis

    PubMed Central

    Marcinkowska, Michalina; Drwęska-Matelska, Natalia; Michalak, Michał; Horst-Sikorska, Wanda; Słomski, Ryszard

    2016-01-01

    Introduction Polymorphism in the promoter region of collagen type 1α (COL1A1) +1245G/T (Sp1, rs1800012) was in some studies shown to be relevant for bone mineral density (BMD) and low-energy fracture prediction. The aim of the study was to confirm this finding in a group of postmenopausal women diagnosed with osteoporosis. Material and methods We investigated 311 Caucasian women (mean age: 65.2 ±9.39 years) either after low-energy fractures (regardless of the location) or meeting World Health Organization (WHO) criteria for osteoporosis. All patients underwent clinical examination in order to exclude secondary osteoporosis; hip and lumbar spine DEXA was performed (Lunar). The three genotypes of Sp1 polymorphism were determined by RFLP (restriction fragment length polymorphism). Results Distribution of COL1A1 genotypes (SS/Ss/ss) agreed with Hardy-Weinberg equilibrium. No relation between COL1A1 genotypes and hip/L1-L4 BMD was found. Fractures were reported in 26.3% of women. Prevalence of low-energy fractures, regardless of the type, was 50.0% in ss genotype carriers, 26.4% in SS homozygotes and 23.7% in Ss heterozygotes. There was no statistically significant recessive or dominant effect of any Sp1 genotype on fracture prevalence (p = 0.613). Conclusions We failed to observe that COL1A1 Sp 1 genotypes contribute to BMD determination or are associated with prevalent low-energy fractures in a Polish cohort of postmenopausal osteoporotic women. PMID:27186171

  10. COL8A1 rs13095226 polymorphism shows no association with neovascular age-related macular degeneration or polypoidal choroidal vasculopathy in Chinese subjects

    PubMed Central

    Yu, Yang; Huang, Lvzhen; Wang, Bin; Zhang, Chunfang; Bai, Yujing; Li, Xiaoxin

    2015-01-01

    Purpose: Age-related macular degeneration (AMD) is the main cause of visual impairment and legal blindness in older individuals. COL8A1 rs13095226 variants have recently been implicated associated with neovascular age-related macular degeneration (nAMD) and Polypoidal Choroidal Vasculopathy (PCV) in American studies. The aim of this study was to investigate the association between the COL8A1 rs13095226 Polymorphism and neovascular age-related macular degeneration (nAMD) and polypoidal choroidal vasculopathy (PCV) in Chinese people. Methods: 900 Chinese subjects-300 cases with nAMD, 300 cases with PCV and 300 controls, were enrolled in a cross-sectional observational study. The diagnoses of nAMD and PCV were confirmed by Fundus photography, Fluorescence Fundus Angiography (FFA) and Indocyanine Green Angiography (ICGA). Genomic DNA was extracted from venous blood leukocytes and genotypes of rs13095226 were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Differences in allele distribution between cases and controls were tested by chi-square tests, with age and gender adjusted by logistic regression analysis. Result: The COL8A1 rs13095226 polymorphism was not statistically significantly different from the nAMD or PCV to the normal controls (P>0.05) in Chinese Population. The association remained insignificant after adjustmentfor age and gender differences (P>0.05). Conclusions: This case-control study indicated that the COL8A1 rs13095226 polymorphism is not associated with nAMD or PCV, which suggesting this gene maybe not a susceptibility gene locus for nAMD or PCV in Chinese subjects. PMID:26617902

  11. Fuyuan Decoction Enhances SOX9 and COL2A1 Expression and Smad2/3 Phosphorylation in IL-1β-Activated Chondrocytes

    PubMed Central

    Zhang, Yudi; Li, Rongheng; Zhong, Yu; Zhang, Sihan; Zhou, Lingyun; Shang, Shike

    2015-01-01

    Fuyuan Decoction (FYD), a herbal formula in China, has been widely used for osteoarthritis (OA) treatment. Herein, we determined the effects of FYD on the expression of transcription factor SOX9 and its target gene collagen type II, alpha 1 (COL2A1) as well as the activation of Smad2/3 in interleukin- (IL-) 1β-stimulated SW1353 chondrosarcoma cells. Serum-derived FYD (FYD-CS) was prepared to treat SW1353 cells with or without SB431542, a TGF-β1 receptor inhibitor. Cell cycle progression was tested by flow cytometry. The expression of SOX9 and COL2A1 and the activation of Smad2/3 (p-Smad2/3) were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and/or western blot. The results showed that, after treatment, FYD-CS, while inducing S-phase cell cycle arrest, enhanced cell proliferation and protected the cells against IL-1β- and/or SB431542-induced cell growth inhibition. Furthermore, FYD-CS reversed the decreased expression of COL2A1 and SOX9 induced by IL-1β and SB431542 and blocked the decreased phosphorylation of Smad2/3 induced by IL-1β alone or in combination with SB431542. Our results suggest that FYD promotes COL2A1 and SOX9 expression as well as Smad2/3 activation in IL-1β-induced chondrocytes, thus benefiting cell survival. PMID:26770254

  12. Congenital Myasthenic Syndrome Type 19 Is Caused by Mutations in COL13A1, Encoding the Atypical Non-fibrillar Collagen Type XIII α1 Chain

    PubMed Central

    Logan, Clare V.; Cossins, Judith; Rodríguez Cruz, Pedro M.; Parry, David A.; Maxwell, Susan; Martínez-Martínez, Pilar; Riepsaame, Joey; Abdelhamed, Zakia A.; Lake, Alice V.R.; Moran, Maria; Robb, Stephanie; Chow, Gabriel; Sewry, Caroline; Hopkins, Philip M.; Sheridan, Eamonn; Jayawant, Sandeep; Palace, Jacqueline; Johnson, Colin A.; Beeson, David

    2015-01-01

    The neuromuscular junction (NMJ) consists of a tripartite synapse with a presynaptic nerve terminal, Schwann cells that ensheathe the terminal bouton, and a highly specialized postsynaptic membrane. Synaptic structural integrity is crucial for efficient signal transmission. Congenital myasthenic syndromes (CMSs) are a heterogeneous group of inherited disorders that result from impaired neuromuscular transmission, caused by mutations in genes encoding proteins that are involved in synaptic transmission and in forming and maintaining the structural integrity of NMJs. To identify further causes of CMSs, we performed whole-exome sequencing (WES) in families without an identified mutation in known CMS-associated genes. In two families affected by a previously undefined CMS, we identified homozygous loss-of-function mutations in COL13A1, which encodes the alpha chain of an atypical non-fibrillar collagen with a single transmembrane domain. COL13A1 localized to the human muscle motor endplate. Using CRISPR-Cas9 genome editing, modeling of the COL13A1 c.1171delG (p.Leu392Sfs∗71) frameshift mutation in the C2C12 cell line reduced acetylcholine receptor (AChR) clustering during myotube differentiation. This highlights the crucial role of collagen XIII in the formation and maintenance of the NMJ. Our results therefore delineate a myasthenic disorder that is caused by loss-of-function mutations in COL13A1, encoding a protein involved in organization of the NMJ, and emphasize the importance of appropriate symptomatic treatment for these individuals. PMID:26626625

  13. Congenital Myasthenic Syndrome Type 19 Is Caused by Mutations in COL13A1, Encoding the Atypical Non-fibrillar Collagen Type XIII α1 Chain.

    PubMed

    Logan, Clare V; Cossins, Judith; Rodríguez Cruz, Pedro M; Parry, David A; Maxwell, Susan; Martínez-Martínez, Pilar; Riepsaame, Joey; Abdelhamed, Zakia A; Lake, Alice V R; Moran, Maria; Robb, Stephanie; Chow, Gabriel; Sewry, Caroline; Hopkins, Philip M; Sheridan, Eamonn; Jayawant, Sandeep; Palace, Jacqueline; Johnson, Colin A; Beeson, David

    2015-12-01

    The neuromuscular junction (NMJ) consists of a tripartite synapse with a presynaptic nerve terminal, Schwann cells that ensheathe the terminal bouton, and a highly specialized postsynaptic membrane. Synaptic structural integrity is crucial for efficient signal transmission. Congenital myasthenic syndromes (CMSs) are a heterogeneous group of inherited disorders that result from impaired neuromuscular transmission, caused by mutations in genes encoding proteins that are involved in synaptic transmission and in forming and maintaining the structural integrity of NMJs. To identify further causes of CMSs, we performed whole-exome sequencing (WES) in families without an identified mutation in known CMS-associated genes. In two families affected by a previously undefined CMS, we identified homozygous loss-of-function mutations in COL13A1, which encodes the alpha chain of an atypical non-fibrillar collagen with a single transmembrane domain. COL13A1 localized to the human muscle motor endplate. Using CRISPR-Cas9 genome editing, modeling of the COL13A1 c.1171delG (p.Leu392Sfs(∗)71) frameshift mutation in the C2C12 cell line reduced acetylcholine receptor (AChR) clustering during myotube differentiation. This highlights the crucial role of collagen XIII in the formation and maintenance of the NMJ. Our results therefore delineate a myasthenic disorder that is caused by loss-of-function mutations in COL13A1, encoding a protein involved in organization of the NMJ, and emphasize the importance of appropriate symptomatic treatment for these individuals. PMID:26626625

  14. Expression profile of COL2A1 and the pseudogene SLC6A10P predicts tumor recurrence in high-grade serous ovarian cancer.

    PubMed

    Ganapathi, Mahrukh K; Jones, Wendell D; Sehouli, Jalid; Michener, Chad M; Braicu, Ioana E; Norris, Eric J; Biscotti, Charles V; Vaziri, Susan A J; Ganapathi, Ram N

    2016-02-01

    Tumor recurrence, following initial response to adjuvant chemotherapy, is a major problem in women with high-grade serous ovarian cancer (HGSOC). Microarray analysis of primary tumors has identified genes that may be useful in risk stratification/overall survival, but are of limited value in predicting the >70% rate for tumor recurrence. In this study, we performed RNA-Seq analysis of primary and recurrent HGSOC to first identify unique differentially expressed genes. From this dataset, we selected 21 archetypical coding genes and one noncoding RNA, based on statistically significant differences in their expression profile between tumors, for validation by qPCR in a larger cohort of 110 ovarian tumors (71 primary and 39 recurrent) and for testing association of specific genes with time-to-recurrence (TTR). Kaplan-Meier tests revealed that high expression of collagen type II, alpha 1 (COL2A1) was associated with delayed TTR (HR = 0.47, 95% CI: 0.27-0.82, p = 0.008), whereas low expression of the pseudogene, solute carrier family 6 member 10 (SLC6A10P), was associated with longer TTR (HR = 0.53, 95% CI: 0.30-0.93, p = 0.027). Notably, TTR was significantly delayed for tumors that simultaneously highly expressed COL2A1 and lowly expressed SLC6A10P (HR = 0.21, 95% CI: 0.082-0.54, p = 0.0011), an estimated median of 95 months as compared to an estimated median of 16 months for subjects expressing other levels of COL2A1 and SLC6A10P. Thus, evaluating expression levels of COL2A1 and SLC6A10P at primary surgery could be beneficial for clinically managing recurrence of HGSOC. PMID:26311224

  15. A familial case of achondrogenesis type II caused by a dominant COL2A1 mutation and "patchy" expression in the mosaic father.

    PubMed

    Forzano, F; Lituania, M; Viassolo, A; Superti-Furga, V; Wildhardt, G; Zabel, B; Faravelli, F

    2007-12-01

    Achondrogenesis type II (ACG2) is the most severe disorder that can be produced by dominant mutations in COL2A1. We report on four pregnancies of an apparently healthy, nonconsanguineous young couple. The father had scoliosis as a child, and has slight body disproportion with short trunk. The first child was born at 32 weeks and died neonatally. In the second pregnancy, short limbs and fetal hygroma were noted on ultrasound at 17 weeks' gestation. Similar findings were observed in the third fetus. Clinical, radiological, and histological evaluation of the fetuses after termination of the pregnancies showed findings consistent with ACG2. Molecular analysis of genomic DNA extracted from amniotic cells of the second and third fetuses revealed heterozygosity for a 10370G > T missense mutation (G346V) in the COL2A1 gene. This mutation was also found in the father, as a mosaic. The couple had a fourth pregnancy, and at 11 weeks fetal hydrops with a septated cystic hygroma were obvious. DNA from CVS demonstrated the same COL2A1 mutation. PMID:17994563

  16. Vascular Ehlers-Danlos Syndrome in siblings with biallelic COL3A1 sequence variants and marked clinical variability in the extended family.

    PubMed

    Jørgensen, Agnete; Fagerheim, Toril; Rand-Hendriksen, Svend; Lunde, Per I; Vorren, Torgrim O; Pepin, Melanie G; Leistritz, Dru F; Byers, Peter H

    2015-06-01

    Vascular Ehlers-Danlos Syndrome (vEDS), also known as EDS type IV, is considered to be an autosomal dominant disorder caused by sequence variants in COL3A1, which encodes the chains of type III procollagen. We identified a family in which there was marked clinical variation with the earliest death due to extensive aortic dissection at age 15 years and other family members in their eighties with no complications. The proband was born with right-sided clubfoot but was otherwise healthy until he died unexpectedly at 15 years. His sister, in addition to signs consistent with vascular EDS, had bilateral frontal and parietal polymicrogyria. The proband and his sister each had two COL3A1 sequence variants, c.1786C>T, p.(Arg596*) in exon 26 and c.3851G>A, p.(Gly1284Glu) in exon 50 on different alleles. Cells from the compound heterozygote produced a reduced amount of type III procollagen, all the chains of which had abnormal electrophoretic mobility. Biallelic sequence variants have a significantly worse outcome than heterozygous variants for either null mutations or missense mutations, and frontoparietal polymicrogyria may be an added phenotype feature. This genetic constellation provides a very rare explanation for marked intrafamilial clinical variation due to sequence variants in COL3A1. PMID:25205403

  17. Mutation in a gene for type I procollagen (COL1A2) in a woman with postmenopausal osteoporosis: Evidence for phenotypic and genotypic overlap with mild osteogenesis imperfecta

    SciTech Connect

    Spotila, L.D.; Constantinou, C.D.; Sereda, L.; Ganguly, A.; Prockop, D.J. ); Riggs, B.L. )

    1991-06-15

    Mutations in the two genes for type I collagen (COL1A1 or COL1A2) cause osteogenesis imperfecta (OI), a heritable disease characterized by moderate to extreme brittleness of bone early in life. Here, the authors show that a 52-year-old post menopausal woman with severe osteopenia and a compression fracture of a thoracic vertebra had a mutation in the gene for the {alpha}2(I) chain of type I collagen (COL1A2) similar to mutations that cause OI. cDNA was prepared from the woman's skin fibroblast RNA and assayed for the presence of a mutation by treating DNA heteroduplexes with carbodiimide. The results indicated a sequence variation in the region encoding amino acid residues 660-667 of the {alpha}2(I) chain. Further analysis demonstrated a single-base mutation that caused a serine-for-glycine substitution at position 661 of the {alpha}2(I) triple-helical domain. The substitution produced posttranslational overmodification of the collagen triple helix, as is seen with most glycine substitutions that cause OI. The patient had a history of five previous fractures, slightly blue sclerae, and slight hearing loss. Therefore, the results suggest that there may be phenotypic and genotypic overlap between mild osteogenesis imperfecta and postmenopausal osteoporosis, and that a subset of women with postmenopausal osteoporosis may have mutations in the genes for type I procollagen.

  18. Endoplasmic reticulum stress-mediated apoptosis contributes to a skeletal dysplasia resembling platyspondylic lethal skeletal dysplasia, Torrance type, in a novel Col2a1 mutant mouse line.

    PubMed

    Kimura, Makoto; Ichimura, Satoki; Sasaki, Kuniaki; Masuya, Hiroshi; Suzuki, Tomohiro; Wakana, Shigeharu; Ikegawa, Shiro; Furuichi, Tatsuya

    In humans, mutations in the COL2A1 gene encoding the α1(II) chain of type II collagen, create many clinical phenotypes collectively termed type II collagenopathies. However, the mechanisms generating this diversity remain to be determined. Here we identified a novel Col2a1 mutant mouse line by screening a large-scale N-ethyl-N-nitrosourea mutant mouse library. This mutant possessed a p.Tyr1391Ser missense mutation in the C-propeptide coding region, and this mutation was located in positions corresponding to the human COL2A1 mutation responsible for platyspondylic lethal skeletal dysplasia, Torrance type (PLSD-T). As expected, p.Tyr1391Ser homozygotes exhibited lethal skeletal dysplasias resembling PLSD-T, including extremely short limbs and severe dysplasia of the spine and pelvis. The secretion of the mutant proteins into the extracellular space was disrupted, accompanied by an abnormally expanded endoplasmic reticulum (ER) and the up-regulation of ER stress-related genes in chondrocytes. Chondrocyte apoptosis was severely induced in the growth plate of the homozygotes. These findings strongly suggest that ER stress-mediated apoptosis caused by the accumulated mutant proteins in ER contributes to skeletal dysplasia in Co12a1 mutant mice and PLSD-T patients. PMID:26545783

  19. Vascular Ehlers–Danlos Syndrome in siblings with biallelic COL3A1 sequence variants and marked clinical variability in the extended family

    PubMed Central

    Jørgensen, Agnete; Fagerheim, Toril; Rand-Hendriksen, Svend; Lunde, Per I; Vorren, Torgrim O; Pepin, Melanie G; Leistritz, Dru F; Byers, Peter H

    2015-01-01

    Vascular Ehlers–Danlos Syndrome (vEDS), also known as EDS type IV, is considered to be an autosomal dominant disorder caused by sequence variants in COL3A1, which encodes the chains of type III procollagen. We identified a family in which there was marked clinical variation with the earliest death due to extensive aortic dissection at age 15 years and other family members in their eighties with no complications. The proband was born with right-sided clubfoot but was otherwise healthy until he died unexpectedly at 15 years. His sister, in addition to signs consistent with vascular EDS, had bilateral frontal and parietal polymicrogyria. The proband and his sister each had two COL3A1 sequence variants, c.1786C>T, p.(Arg596*) in exon 26 and c.3851G>A, p.(Gly1284Glu) in exon 50 on different alleles. Cells from the compound heterozygote produced a reduced amount of type III procollagen, all the chains of which had abnormal electrophoretic mobility. Biallelic sequence variants have a significantly worse outcome than heterozygous variants for either null mutations or missense mutations, and frontoparietal polymicrogyria may be an added phenotype feature. This genetic constellation provides a very rare explanation for marked intrafamilial clinical variation due to sequence variants in COL3A1. PMID:25205403

  20. Null alleles of the COL5A1 gene of type V collagen are a cause of the classical forms of Ehlers-Danlos syndrome (types I and II).

    PubMed Central

    Schwarze, U; Atkinson, M; Hoffman, G G; Greenspan, D S; Byers, P H

    2000-01-01

    Ehlers-Danlos syndrome (EDS) types I and II, which comprise the classical variety, are well characterized from the clinical perspective, but it has been difficult to identify the molecular basis of the disorder in the majority of affected individuals. Several explanations for this failure to detect mutations have been proposed, including genetic heterogeneity, failure of allele expression, and technical difficulties. Genetic heterogeneity has been confirmed as an explanation for such failure, since causative mutations have been identified in the COL5A1, COL5A2, and tenascin X genes and since they have been inferred in the COL1A2 gene. Nonetheless, in the majority of families with autosomal dominant inheritance of EDS, there appears to be linkage to loci that contain the COL5A1 or COL5A2 genes. To determine whether allele-product instability could explain failure to identify some mutations, we analyzed polymorphic variants in the COL5A1 gene in 16 individuals, and we examined mRNA for the expression of both alleles and for alterations in splicing. We found a splice-site mutation in a single individual, and we determined that, in six individuals, the mRNA from one COL5A1 allele either was not expressed or was very unstable. We identified small insertions or deletions in five of these cell strains, but we could not identify the mutation in the sixth individual. Thus, although as many as one-half of the mutations that give rise to EDS types I and II are likely to lie in the COL5A1 gene, a significant portion of them result in very low levels of mRNA from the mutant allele, as a consequence of nonsense-mediated mRNA decay. PMID:10796876

  1. La pseudarthrose du col fémoral traitée par prothèse totale de la hanche: à propos de 15 cas

    PubMed Central

    Chagou, Aniss; Bassir, Réda Allah; Rhanim, Abdelkarim; Lahlou, Abdou; Bardouni, Ahmed; Mahfoud, Moustapha; Saleh Berrada, Mohammed; El Yaacoubi, Moradh

    2014-01-01

    La pseudarthrose du col fémoral est une complication redoutée de la facture du col fémoral, elle est due soit à une négligence thérapeutique, soit à une ostéosynthèse imparfaite. Plusieurs facteurs sont incriminés dans sa genèse, l’âge, les caractéristiques de la fracture, l’état de la tête fémorale, et une ostéosynthèse non solide. Le diagnostic des pseudarthroses est essentiellement radiologique. Le traitement reste encore difficile et mal codifié, les limites entre traitement conservateur et arthroplastie de la hanche ne sont pas encore bien définies. Nous rapportons une série de 15 cas de pseudarthrose du col fémoral traités par arthroplastie de la hanche au service de traumato-orthopédie du Centre Hospitalier Universitaire de Rabat de 2008 à 2014 soit un recul de 40 mois en moyenne. Nos patients ont bénéficié d'une évaluation clinique et radiologique. L’âge de nos patients varie entre 48 et 81 ans, avec une moyenne de 69,2 ans. 85% d'entre eux sont âgés de plus de 60 ans. Nous avons dans notre série une prédominance féminine, soit 8 femmes pour 7 hommes. La négligence thérapeutique est la cause de la majorité des pseudarthroses du col du fémur traitées dans notre série. Par ailleurs, nous avons utilisé exclusivement la voie d'abord postéro externe de Moore. Nous avons mis en place une prothèse totale de couple polyéthylène-métal chez tous les patients, ces prothèses étaient cimentées chez 12 patients. Le résultat fonctionnel a été coté selon la classification de Merle d'Aubigné. Nos résultats ont été jugés bons selon cette cotation. L'arthroplastie totale de la hanche a une place importante dans le traitement des pseudarthroses du col fémoral et peut donner des résultats satisfaisants en permettant de récupérer une hanche mobile et indolore. PMID:25667720

  2. SIRT1 deacetylates RFX5 and antagonizes repression of collagen type I (COL1A2) transcription in smooth muscle cells

    SciTech Connect

    Xia, Jun; Wu, Xiaoyan; Yang, Yuyu; Zhao, Yuhao; Fang, Mingming; Xie, Weiping; Wang, Hong; Xu, Yong

    2012-11-16

    Highlights: Black-Right-Pointing-Pointer SIRT1 interacts with and deacetylates RFX5. Black-Right-Pointing-Pointer SIRT1 activation attenuates whereas SIRT1 inhibition enhances collagen repression by RFX5 in vascular smooth muscle cells. Black-Right-Pointing-Pointer SIRT1 promotes cytoplasmic localization and proteasomal degradation of RFX5 and cripples promoter recruitment of RFX5. Black-Right-Pointing-Pointer IFN-{gamma} represses SIRT1 expression in vascular smooth muscle cells. Black-Right-Pointing-Pointer SIRT1 agonist alleviates collagen repression by IFN-{gamma} in vascular smooth muscle cells. -- Abstract: Decreased expression of collagen by vascular smooth muscle cells (SMCs) within the atherosclerotic plaque contributes to the thinning of the fibrous cap and poses a great threat to plaque rupture. Elucidation of the mechanism underlying repressed collagen type I (COL1A2) gene would potentially provide novel solutions that can prevent rupture-induced complications. We have previously shown that regulatory factor for X-box (RFX5) binds to the COL1A2 transcription start site and represses its transcription. Here we report that SIRT1, an NAD-dependent, class III deacetylase, forms a complex with RFX5. Over-expression of SIRT1 or NAMPT, which synthesizes NAD+ to activate SIRT1, or treatment with the SIRT1 agonist resveratrol decreases RFX5 acetylation and disrupts repression of the COL1A2 promoter activity by RFX5. On the contrary, knockdown of SIRT1 or treatment with SIRT1 inhibitors induces RFX5 acetylation and enhances the repression of collagen transcription. SIRT1 antagonizes RFX5 activity by promoting its nuclear expulsion and proteasomal degradation hence dampening its binding to the COL1A2 promoter. The pro-inflammatory cytokine IFN-{gamma} represses COL1A2 transcription by down-regulating SIRT1 expression in SMCs. Therefore, our data have identified as novel pathway whereby SIRT1 maintains collagen synthesis in SMCs by modulating RFX5 activity.

  3. Prevalence of ColV Plasmid-Linked Genes and In Vivo Pathogenicity of Avian Strains of Escherichia coli.

    PubMed

    de Oliveira, Aline Luísa; Rocha, Débora Assumpção; Finkler, Fabrine; de Moraes, Lucas Brunelli; Barbieri, Nicolle Lima; Pavanelo, Daniel Brisotto; Winkler, Cristina; Grassotti, Tiela Trapp; de Brito, Kelly Cristina Tagliari; de Brito, Benito Guimarães; Horn, Fabiana

    2015-08-01

    Avian pathogenic Escherichia coli (APEC) causes extraintestinal infections in birds, leading to an increase in the cost of poultry production. The ColV plasmid-linked genes iroN, ompT, hlyF, iss, and iutA have previously been suggested to be predictors of the virulence of APEC. In this research, we analyzed the frequencies of these genes in a Brazilian collection of E. coli isolated from birds with colibacillosis (APEC) and from apparently healthy birds (avian fecal [A(fecal)]), as well as from the litter of poultry houses of apparently healthy flocks (avian litter [A(litter)]). All the isolates that harbored ompT also harbored hlyF, so they were considered as one trait for statistical analysis. The relationship between in vivo virulence in 1-day-old chicks, expressed as a pathogenicity score, and the number of genes in each isolate showed that isolates with less than two of the four genes were rarely pathogenic, while most pathogenic isolates contained two or more genes. Nevertheless, about half of the nonpathogenic isolates also harbored two or more genes, in agreement with previous observations that commensal E. coli isolates from the birds' microbiota can serve as a reservoir of virulence genes. Thus, the pentaplex polymerase chain reaction can be used to indicate that a strain carrying none or only one gene would be nonpathogenic, but it cannot be used to indicate that a strain with two to four genes would be an APEC. Isolates allocated to phylogenetic group B2, which is frequently associated with extraintestinal infections, had the highest pathogenicity scores, while isolates allocated to group B1 had the lowest. PMID:26258262

  4. RFX family proteins differentially interact with HDACs to repress collagen alpha 2(I) gene (COL1A2) expression

    PubMed Central

    Xu, Y.; Sengupta, P.K.; Seto, E.; Smith, B.D.

    2006-01-01

    Our studies indicate that regulatory factor for X-box (RFX) family proteins repress collagen alpha2(I) gene (COL1A2) expression (1,2). In the present investigation, we examine the mechanism(s) underlying the repression of collagen gene by RFX proteins. Two members of the RFX family, RFX1 and RFX5, associate with distinct sets of co-repressors on the collagen transcription start site in vitro. RFX5 specifically interacts with histone deacetylase 2 (HDAC2) and the mammalian transcriptional repressor (mSin3B) whereas RFX1 preferably interacts with HDAC1 and mSin3A. HDAC2 cooperates with RFX5 to down-regulate collagen promoter activity while HDAC1 enhances inhibition of collagen promoter activity by RFX1. IFN-γ promotes the recruitment of RFX5/HDAC2/mSin3B to the collagen transcription start site but decreases the occupancy by RFX1/mSin3A as manifested by chromatin immunoprecipitation (ChIP) assay. RFX1 binds to methylated collagen sequence with much higher affinity than unmethylated sequence, recruiting more HDAC1 and mSin3A. The DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (aza-dC), that inhibits DNA methylation, reduces RFX1/HDAC1 binding to the collagen transcription start site in ChIP assays. Finally, both RFX1 and RFX5 are acetylated in vivo. TSA stimulates the acetylation of RFX proteins and activates the collagen promoter activity. Collectively, our data strongly indicate two separate pathways for RFX proteins to repress collagen gene expression: one for RFX5/HDAC2 in IFN-γ mediated repression, the other for RFX1/HDAC1 in methylation mediated collagen silencing. PMID:16464847

  5. Regulation of COL1A1 expression in type I collagen producing tissues: identification of a 49 base pair region which is required for transgene expression in bone of transgenic mice

    NASA Technical Reports Server (NTRS)

    Bedalov, A.; Salvatori, R.; Dodig, M.; Kronenberg, M. S.; Kapural, B.; Bogdanovic, Z.; Kream, B. E.; Woody, C. O.; Clark, S. H.; Mack, K.; Rowe, D. W. (Principal Investigator)

    1995-01-01

    Previous deletion studies using a series of COL1A1-CAT fusion genes have indicated that the 625 bp region of the COL1A1 upstream promoter between -2295 and -1670 bp is required for high levels of expression in bone, tendon, and skin of transgenic mice. To further define the important sequences within this region, a new series of deletion constructs extending to -1997, -1794, -1763, and -1719 bp has been analyzed in transgenic mice. Transgene activity, determined by measuring CAT activity in tissue extracts of 6- to 8-day-old transgenic mouse calvariae, remains high for all the new deletion constructs and drops to undetectable levels in calvariae containing the -1670 bp construct. These results indicate that the 49 bp region of the COL1A1 promoter between -1719 and -1670 bp is required for high COL1A1 expression in bone. Although deletion of the same region caused a substantial reduction of promoter activity in tail tendon, the construct extending to -1670 bp is still expressed in this tissue. However, further deletion of the promoter to -944 bp abolished activity in tendon. Gel mobility shift studies identified a protein in calvarial nuclear extracts that is not found in tendon nuclear extracts, which binds within this 49 bp region. Our study has delineated sequences in the COL1A1 promoter required for expression of the COL1A1 gene in high type I collagen-producing tissues, and suggests that different cis elements control expression of the COL1A1 gene in bone and tendon.

  6. PCR-SSCP analysis of the type VII collagen gene (COL7A1): Detection of a point mutation in five patients

    SciTech Connect

    Dunnil, M.G.S.; Richards, A.J.; Pope, F.M.

    1994-09-01

    Type VII collagen is the major component of anchoring fibrils, structures which extend below the lamina densa of the epidermal basement membrane in stratified squamous epithelia. Genetic linkage studies and two mutation reports have implicated the type VII collagen gene, COL7A1, in dystrophic epidermolysis bullosa (DEB), an inherited disorder characterized by blistering and scarring of the skin and mucous membranes after minor trauma. We have used PCR-SSCP of genomic DNA to screen exons of COL7A1 for mutations in recessive DEB patients. Band mobility shifts were detected in exon FN4-B in five patients. Sequencing revealed a C to T transition changing a codon for arginine into a stop codon, homozygous in two related patients and heterozygous in the others. We are currently searching for a second mutation in these three heterozygous patients who are presumably genetic compounds. Screening for an informative Xho I restriction site altered by the mutation showed parental heterozygosity but no evidence for the mutation in 50 normal chromosomes. Segregation of COL7A1 markers in these patients suggests that the mutation has arisen independently in at least two of our families. The premature stop mutation in the 5{prime} end of the gene predicts a severely shortened collagen VII molecule. The homozygote formation of anchoring fibrils would be impaired providing an explanation at the molecular level for the ultrastructural findings of reduced numbers or absence of anchoring fibrils in this disease. In conclusion, these data strongly suggest that this novel premature stop mutation is the cause of DEB in the homozygotes and contributes to the disease in the other patients. The important role of anchoring fibrils in dermal-epidermal adhesion is also underlined.

  7. Cloning of the human type XVII collagen gene (COL17A1), and detection of novel mutations in generalized atrophic benign epidermolysis bullosa

    SciTech Connect

    Gatalica, B.; Pulkkinen, L.; Li, K.

    1997-02-01

    Generalized atrophic benign epidermolysis bullosa (GABEB) is a nonlethal variant of junctional epidermolysis bullosa (JEB). Previous findings have suggested that type XVII collagen is the candidate gene for mutations in this disease. We now have cloned the entire human type XVII collagen gene (COL17A1) and have elucidated its intron-exon organization. The gene comprises 56 distinct exons, which span {approximately}52 kb of the genome, on the long arm of chromosome 10. It encodes a polypeptide, the {alpha}1(XVII) chain, consisting of an intracellular globular domain, a transmembrane segment, and an extracellular domain that contains 15 separate collagenous subdomains, the largest consisting of 242 amino acids. We also have developed a strategy to identify mutations in COL17A1 by use of PCR amplification of genomic DNA, using primers placed on the flanking introns. The PCR products are scanned for sequence variants by heteroduplex analysis using conformation-sensitive gel electrophoresis and then are subjected to direct automated sequencing. We have identified several intragenic polymorphisms in COL17A1, as well as mutations, in both alleles, in two Finnish families with GABEB. The probands in both families showed negative immunofluorescence staining with an anti-type XVII collagen antibody. In one family, the proband was homozygous for a 5-bp deletion, 2944del5, which resulted in frameshift and a premature termination codon of translation. The proband in the other family was a compound heterozygote, with one allele containing the 2944del5 mutation and the other containing a nonsense mutation, Q1023X. These results expand the mutation database in different variants of JEB, and they attest to the functional importance of type XVII collagen as a transmembrane component of the hemidesmosomes at the dermal/epidermal junction. 48 refs., 9 figs., 3 tabs.

  8. Double replacement: strategy for efficient introduction of subtle mutations into the murine Col1a-1 gene by homologous recombination in embryonic stem cells.

    PubMed Central

    Wu, H; Liu, X; Jaenisch, R

    1994-01-01

    A subtle mutation that rendered type I collagen resistant to mammalian collagenase has been introduced into the murine Col1a-1 (recently redesignated Cola-1) gene by homologous recombination in embryonic stem (ES) cells. Initially, a "hit and run" procedure was used. Since two steps were required for introducing each mutation and more than one mutation was to be introduced in the same genomic region independently, we have developed a streamlined procedure that involves two sequential replacement-type homologous recombination events. In the first step, an internal deletion was introduced into the Col1a-1 locus along with the positive and negative selectable markers, neo and tk, to mark the region of interest. G418-resistant homologous recombinants were isolated and used in the second step in which the deleted Col1a-1 allele was replaced with a construct containing the desired mutation. Homologous recombinants containing the mutation were identified among the Tk- ES clones after selection with FIAU [1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil (called fialuridine)]. Approximately 10% of such clones contained the desired mutation. The double replacement procedure greatly reduces the time and amount of work required to introduce mutations independently into the same or closely linked regions. Once the homologous recombinants derived from the first step are established, the introduction of other mutations into the deleted region becomes a one-step procedure. For X number of introduced mutations, 2X selections are required with the "hit and run" approach, but only X + 1 are required with the double-replacement method. This innovative procedure could be very useful in studies of gene structure and function as well as gene expression and regulation. Images PMID:8146196

  9. Targeting the annulus fibrosus of the intervertebral disc: Col1a2-Cre(ER)T mice show specific activity of Cre recombinase in the outer annulus fibrosus.

    PubMed

    Bedore, Jake; Quesnel, Katherine; Quinonez, Diana; Séguin, Cheryle A; Leask, Andrew

    2016-06-01

    Degeneration of the intervertebral disc (IVD) is a major underlying contributor to back pain-the single leading cause of disability worldwide. However, we possess a limited understanding of the etiology underlying IVD degeneration. To date, there are a limited number of mouse models that have been used to target proteins in specific compartments of the IVD to explore their functions in disc development, homeostasis and disease. Furthermore, the majority of reports exploring the composition and function of the outer encapsulating annulus fibrosus (AF) of the IVD have considered it as one tissue, without considering the numerous structural and functional differences existing between the inner and outer AF. In addition, no mouse models have yet been reported that enable specific targeting of genes within the outer AF. In the current report, we discuss these issues and demonstrate the localized activity of Cre recombinase in the IVD of Col1a2-Cre(ER)T;ROSA26mTmG mice possessing a tamoxifen-dependent Cre recombinase driven by a Cola2 promoter and distal enhancer and the mTmG fluorescent reporter. Following tamoxifen injection of 3-week-old Col1a2-Cre(ER)T;ROSA26mTmG mice, we show Cre activity specifically in the outer AF of the IVD, as indicated by expression of the GFP reporter. Thus, Col1a2-Cre(ER)T;ROSA26mTmG mice may prove to be a valuable tool in delineating the function of proteins in this unique compartment of the IVD, and in further exploring the compositional differences between the inner and outer AF in disc homeostasis, aging and disease. PMID:27173473

  10. A Site-Specific Integrated Col2.3GFP Reporter Identifies Osteoblasts Within Mineralized Tissue Formed In Vivo by Human Embryonic Stem Cells.

    PubMed

    Xin, Xiaonan; Jiang, Xi; Wang, Liping; Stover, Mary Louise; Zhan, Shuning; Huang, Jianping; Goldberg, A Jon; Liu, Yongxing; Kuhn, Liisa; Reichenberger, Ernst J; Rowe, David W; Lichtler, Alexander C

    2014-10-01

    The use of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) for study and treatment of bone diseases or traumatic bone injuries requires efficient protocols to differentiate hESCs/iPSCs into cells with osteogenic potential and the ability to isolate differentiated osteoblasts for analysis. We have used zinc finger nuclease technology to deliver a construct containing the Col2.3 promoter driving GFPemerald to the AAVS1 site (referred to as a "safe harbor" site), in human embryonic stem cells (H9Zn2.3GFP), with the goal of marking the cells that have become differentiated osteoblasts. In teratomas formed using these cells, we identified green fluorescent protein (GFP)-positive cells specifically associated with in vivo bone formation. We also differentiated the cells into a mesenchymal stem cell population with osteogenic potential and implanted them into a mouse calvarial defect model. We observed GFP-positive cells associated with alizarin complexone-labeled newly formed bone surfaces. The cells were alkaline phosphatase-positive, and immunohistochemistry with human specific bone sialoprotein (BSP) antibody indicates that the GFP-positive cells are also associated with the human BSP-containing matrix, demonstrating that the Col2.3GFP construct marks cells in the osteoblast lineage. Single-cell cloning generated a 100% Col2.3GFP-positive cell population, as demonstrated by fluorescence in situ hybridization using a GFP probe. The karyotype was normal, and pluripotency was demonstrated by Tra1-60 immunostaining, pluripotent low density reverse transcription-polymerase chain reaction array and embryoid body formation. These cells will be useful to develop optimal osteogenic differentiation protocols and to isolate osteoblasts from normal and diseased iPSCs for analysis. PMID:25122686

  11. A Site-Specific Integrated Col2.3GFP Reporter Identifies Osteoblasts Within Mineralized Tissue Formed In Vivo by Human Embryonic Stem Cells

    PubMed Central

    Xin, Xiaonan; Jiang, Xi; Wang, Liping; Stover, Mary Louise; Zhan, Shuning; Huang, Jianping; Goldberg, A. Jon; Liu, Yongxing; Kuhn, Liisa; Reichenberger, Ernst J.; Rowe, David W.

    2014-01-01

    The use of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) for study and treatment of bone diseases or traumatic bone injuries requires efficient protocols to differentiate hESCs/iPSCs into cells with osteogenic potential and the ability to isolate differentiated osteoblasts for analysis. We have used zinc finger nuclease technology to deliver a construct containing the Col2.3 promoter driving GFPemerald to the AAVS1 site (referred to as a “safe harbor” site), in human embryonic stem cells (H9Zn2.3GFP), with the goal of marking the cells that have become differentiated osteoblasts. In teratomas formed using these cells, we identified green fluorescent protein (GFP)-positive cells specifically associated with in vivo bone formation. We also differentiated the cells into a mesenchymal stem cell population with osteogenic potential and implanted them into a mouse calvarial defect model. We observed GFP-positive cells associated with alizarin complexone-labeled newly formed bone surfaces. The cells were alkaline phosphatase-positive, and immunohistochemistry with human specific bone sialoprotein (BSP) antibody indicates that the GFP-positive cells are also associated with the human BSP-containing matrix, demonstrating that the Col2.3GFP construct marks cells in the osteoblast lineage. Single-cell cloning generated a 100% Col2.3GFP-positive cell population, as demonstrated by fluorescence in situ hybridization using a GFP probe. The karyotype was normal, and pluripotency was demonstrated by Tra1-60 immunostaining, pluripotent low density reverse transcription-polymerase chain reaction array and embryoid body formation. These cells will be useful to develop optimal osteogenic differentiation protocols and to isolate osteoblasts from normal and diseased iPSCs for analysis. PMID:25122686

  12. Upregulated Expression of Integrin α1 in Mesangial Cells and Integrin α3 and Vimentin in Podocytes of Col4a3-Null (Alport) Mice

    PubMed Central

    Steenhard, Brooke M.; Vanacore, Roberto; Friedman, David; Zelenchuk, Adrian; Stroganova, Larysa; Isom, Kathryn; St. John, Patricia L.; Hudson, Billy G.; Abrahamson, Dale R.

    2012-01-01

    Alport disease in humans, which usually results in proteinuria and kidney failure, is caused by mutations to the COL4A3, COL4A4, or COL4A5 genes, and absence of collagen α3α4α5(IV) networks found in mature kidney glomerular basement membrane (GBM). The Alport mouse harbors a deletion of the Col4a3 gene, which also results in the lack of GBM collagen α3α4α5(IV). This animal model shares many features with human Alport patients, including the retention of collagen α1α2α1(IV) in GBMs, effacement of podocyte foot processes, gradual loss of glomerular barrier properties, and progression to renal failure. To learn more about the pathogenesis of Alport disease, we undertook a discovery proteomics approach to identify proteins that were differentially expressed in glomeruli purified from Alport and wild-type mouse kidneys. Pairs of cy3- and cy5-labeled extracts from 5-week old Alport and wild-type glomeruli, respectively, underwent 2-dimensional difference gel electrophoresis. Differentially expressed proteins were digested with trypsin and prepared for mass spectrometry, peptide ion mapping/fingerprinting, and protein identification through database searching. The intermediate filament protein, vimentin, was upregulated ∼2.5 fold in Alport glomeruli compared to wild-type. Upregulation was confirmed by quantitative real time RT-PCR of isolated Alport glomeruli (5.4 fold over wild-type), and quantitative confocal immunofluorescence microscopy localized over-expressed vimentin specifically to Alport podocytes. We next hypothesized that increases in vimentin abundance might affect the basement membrane protein receptors, integrins, and screened Alport and wild-type glomeruli for expression of integrins likely to be the main receptors for GBM type IV collagen and laminin. Quantitative immunofluorescence showed an increase in integrin α1 expression in Alport mesangial cells and an increase in integrin α3 in Alport podocytes. We conclude that overexpression of

  13. Variants of COL3A1 are associated with the risk of stroke recurrence and prognosis in the Chinese population: a prospective study.

    PubMed

    Lv, Wenfei; Lin, Yahui; Song, Weihua; Sun, Kai; Yu, Hui; Zhang, Yinhui; Zhang, Channa; Li, Liang; Suo, Miaomiao; Hui, Rutai; Chen, Jingzhou

    2014-06-01

    Type III collagen plays an important role in activating platelets, forming thrombus, and maintaining the mechanical properties of arteries. This study aimed to test the hypothesis that genetic variants of COL3A1 (gene encoding type III collagen) contribute to recurrence and prognosis of stroke. We investigated the associations of three variants (rs2138533, rs11887092, and rs1800255) in the COL3A1 gene with stroke recurrence and prognosis in 1,544 patients with three subtypes of stroke: lacunar infarction (n = 442), atherothrombotic infarction (n = 670), and hemorrhage (n = 432). These associations were evaluated by Kaplan-Meier analysis and Cox regression models. Patients were followed up for 4.5 years. The A allele of rs1800255 in the COL3A1 gene coding region was significantly associated with a reduced risk of stroke recurrence in patients with lacunar infarction (adjusted hazard ratio [HR] 0.58, 95 % confidence interval [CI] 0.36-0.93, P = 0.024), but there was an increased risk of all-cause mortality of atherothrombotic patients (adjusted HR 1.43, 95 % CI 1.01-2.00, P = 0.044). The TT genotype of rs2138533 showed a significantly increased risk of death caused by cardiovascular disease or stroke in lacunar infarct patients (adjusted HR 2.98, 95 % CI 1.27-6.98, P = 0.012), but there was a reduced risk of all-cause mortality for patients with intracerebral hemorrhage (adjusted HR 0.34, 95 % CI 0.12-0.93, P = 0.036). The G allele of rs11887092 increased the risk of stroke recurrence in patients with atherothrombotic stroke (adjusted HR 1.59, 95 % CI 1.04-2.44, P = 0.035). In conclusion, variants of COL3A1 might play a vital role in determining the risk of recurrence and prognosis after stroke. PMID:24664438

  14. Cervical artery dissections and type A aortic dissection in a family with a novel missense COL3A1 mutation of vascular type Ehlers-Danlos syndrome.

    PubMed

    Makrygiannis, Georgios; Loeys, Bart; Defraigne, Jean-Olivier; Sakalihasan, Natzi

    2015-11-01

    Cervical artery dissection (CeAD) is a rare condition. One of the causes is the vascular type of Ehlers-Danlos syndrome (vEDS). A novel missense mutation in COL3A1 was found in a young patient with CeAD as the single manifestation of vEDS. This is a heterozygous c.953G > A mutation in exon 14, disrupting the normal Gly-X-Y repeats of type III procollagen, by converting glycine to aspartic acid. PMID:26497932

  15. The TATA-containing core promoter of the type II collagen gene (COL2A1) is the target of interferon-gamma-mediated inhibition in human chondrocytes: requirement for Stat1 alpha, Jak1 and Jak2.

    PubMed Central

    Osaki, Makoto; Tan, Lujian; Choy, Bob K; Yoshida, Yasuhiro; Cheah, Kathryn S E; Auron, Philip E; Goldring, Mary B

    2003-01-01

    Interferon-gamma (IFN-gamma) inhibits the synthesis of the cartilage-specific extracellular matrix protein type II collagen, and suppresses the expression of the type II collagen gene ( COL2A1 ) at the transcriptional level. To further examine this mechanism, the responses of COL2A1 regulatory sequences to IFN-gamma and the role of components of the Janus kinase/signal transducer and activators of transcription (JAK/STAT) pathway were examined in the immortalized human chondrocyte cell line, C-28/I2. IFN-gamma inhibited the mRNA levels of COL2A1 and aggrecan, but not Sox9, L-Sox5 and Sox6, all of which were expressed by these cells as markers of the differentiated phenotype. IFN-gamma suppressed the expression of luciferase reporter constructs containing sequences of the COL2A1 promoter spanning -6368 to +125 bp in the absence and presence of the intronic enhancer and stimulated activity of the gamma-interferon-activated site (GAS) luciferase reporter vector, associated with induction of Stat1 alpha-binding activity in nuclear extracts. These responses to IFN-gamma were blocked by overexpression of the JAK inhibitor, JAK-binding protein (JAB), or reversed by dominant-negative Stat1 alpha Y701F containing a mutation at Tyr-701, the JAK phosphorylation site. IFN-gamma had no effect on COL2A1 promoter expression in Jak1 (U4A)-, Jak2 (gamma 2A)- and Stat1 alpha (U3A)-deficient cell lines. In the U3A cell line, the response to IFN-gamma was rescued by overexpression of Stat1 alpha, but not by either Stat1 alpha Y701F or Stat1 beta. Functional analysis using deletion constructs showed that the IFN-gamma response was retained in the COL2A1 core promoter region spanning -45 to +11 bp, containing the TATA-box and GC-rich sequences but no Stat1-binding elements. Inhibition of COL2A1 promoter activity by IFN-gamma persisted in the presence of multiple deletions within the -45/+11 bp region. Our results indicate that repression of COL2A1 gene transcription by IFN

  16. Compound heterozygosity for COL7A1 mutations in twins with dystrophic epidermolysis bullosa: A recessive paternal deletion/insertion mutation and a dominant negative maternal glycine substitution result in a severe phenotype

    SciTech Connect

    Christiano, A.M.; Uitto, J.; Anton-Lamprecht, I.; Ebschner, U.; Amano, S.; Burgeson, R.E.

    1996-04-01

    We have previously demonstrated genetic linkage between the type VII collagen gene (COL7A1) and the dominant (DDEB) and recessive (RDEB) forms of dystrophic epidermolysis bullosa (DEB) and have subsequently identified pathogenetic mutations in several families. Mutations in DDEB identified thus far are glycine substitutions in the collagenous domain of COL7A1, while the most severe forms of RDEB result from premature termination codon (PTC) mutations on both alleles. In this study, we performed mutation analysis in the COL7A1 gene in twins who displayed a severe DEB phenotype. Mutational analysis revealed a paternal 2-bp deletion/1-bp insertion in exon 56, designated 5103CC{yields}G, which results in a frameshift and downstream PTC. Analysis of the maternal COL7A1 allele revealed a glycine-to-arginine substitution in exon 91 (G2351R). Careful questioning of the mother revealed that she and her father had a history of shedding of toenails and occasional poorly heating erosions, consistent with a mild form of DDEB. Immunoprecipitation of type VII collagen from fibroblasts of the twins revealed a marked reduction in intracellular protein production, consistent with the drastic reduction in mRNA transcript from the paternal mutant allele, while the majority of polypeptides bearing the glycine substitution appeared to be degraded intracellularly. Thus, the severe RDEB phenotype in the probands results from compound heterozygosity for one glycine substitution and one PTC mutation in COL7A1. 40 refs., 7 figs.

  17. Parental somatic and germ-line mosaicism for a multiexon deletion with unusual endpoints in a type III collagen (COL3A1) allele produces Ehlers-Danlos syndrome type IV in the heterozygous offspring.

    PubMed Central

    Milewicz, D M; Witz, A M; Smith, A C; Manchester, D K; Waldstein, G; Byers, P H

    1993-01-01

    Ehlers-Danlos syndrome (EDS) type IV is a dominantly inherited disorder that results from mutations in the type III collagen gene (COL3A1). We studied the structure of the COL3A1 gene of an individual with EDS type IV and that of her phenotypically normal parents. The proband was heterozygous for a 2-kb deletion in COL3A1, while her father was mosaic for the same deletion in somatic and germ cells. In fibroblasts from the father, approximately two-fifths of the COL3A1 alleles carried the deletion, but only 10% of the COL3A1 alleles in white blood cells were of the mutant species. The deletion in the mutant allele extended from intron 7 into intron 11. There was a 12-bp direct repeat in intron 7 and intron 11, the latter about 60 bp 5' to the junction. At the breakpoint there was a duplication of 10 bp from intron 11 separated by an insertion of 4 bp contained within the duplicated sequence. The father was mosaic for the deletion so that the gene rearrangement occurred during his early embryonic development prior to lineage allocation. These findings suggest that at least some of the deletions seen in human genes may occur during replication, rather than as a consequence of meiotic crossing-over, and that they thus have a risk for recurrence when observed de novo. Images Figure 1 Figure 2 Figure 3 PMID:8317500

  18. Parental somatic and germ-line mosaicism for a multiexon deletion with unusual endpoints in a type III collagen (COL3Al) allele produces ehlers-danlos syndrome type IV in the heterozygous offspring

    SciTech Connect

    McGookey Milewicz, D.; Witz, A.M.; Byers, P.H. ); Smith, A.C.M.; Manchester, D.K.; Waldstein, G. )

    1993-07-01

    Ehlers-Danlos syndrome (EDS) type IV is a dominantly inherited disorder that results from mutation in the type III collagen gene (COL3A1). The authors studied the structure of the COL3A1 gene of an individual with EDS type IV and that of her phenotypically normal parents. The proband was heterozygous for a 2-kb deletion in COL3A1, while her father was mosaic for the same deletion in somatic and germ cells. In fibroblasts from the father, approximately two-fifths of the COL3A1 alleles carried the deletion, but only 10% of the COL3A1 alleles in white blood cells were of the mutant species. The deletion in the mutant allele extended from intron 7 into intron 11. There was a 12-bp direct repeat in intron 7 and intron 11, the latter about 60 bp 5' to the junction. At the breakpoint there was a duplication of 10 bp from intron 11 separated by an insertion of 4 bp contained within the duplicated sequence. The father was mosaic for the deletion so that the gene rearrangement occurred during his early embryonic development prior to lineage allocation. These findings suggest that at least some of the deletions seen in human genes may occur during replication, rather than as a consequence of meiotic crossing-over, and that they thus have a risk for recurrence when observed de novo. 71 refs., 4 figs., 2 tabs.

  19. An Incompletely Penetrant Novel Mutation in COL7A1 Causes Epidermolysis Bullosa Pruriginosa and Dominant Dystrophic Epidermolysis Bullosa Phenotypes in an Extended Kindred

    PubMed Central

    Yang, Catherine S; Lu, Yin; Farhi, Anita; Nelson-Williams, Carol; Kashgarian, Michael; Glusac, Earl J; Lifton, Richard P; Antaya, Richard J; Choate, Keith A

    2012-01-01

    Epidermolysis bullosa pruriginosa (EBP) is a rare subtype of dystrophic epidermolysis bullosa (DEB) characterized by intense pruritus, nodular or lichenoid lesions, and violaceous linear scarring, most prominently on the extensor extremities. Remarkably, identical mutations in COL7A1, which encodes an anchoring fibril protein present at the dermal–epidermal junction, can cause both DEB and EBP with either autosomal dominant or recessive inheritance. We present one family with both dystrophic and pruriginosa phenotypes of epidermolysis bullosa. The proband is a 19-year-old Caucasian woman who initially presented in childhood with lichenoid papules affecting her extensor limbs and intense pruritus consistent with EBP. Her maternal grandmother saw a dermatologist for similar skin lesions that developed without any known triggers at age 47 and mostly resolved spontaneously after approximately 10 years. The proband’s younger brother developed a small crop of pruritic papules on his elbows, dorsal hands, knees, and ankles at age 13. Her second cousin once removed, however, reported a mild blistering disease without pruritus consistent with DEB. Genetic sequencing of the kindred revealed a single dominant novel intron 47 splice site donor G>A mutation, c.4668 + 1 G>A, which we predict leads to exon skipping. Incomplete penetrance is confirmed in her clinically unaffected mother, who carries the same dominant mutation. The wide diversity of clinical phenotypes with one underlying genotype demonstrates that COL7A1 mutations are incompletely penetrant and strongly suggests that other genetic and environmental factors influence clinical presentation. PMID:22515571

  20. Soft-tissue aneurysmal bone cyst with translocation t(17;17)(p13;q21) corresponding to COL1A1 and USP6 loci.

    PubMed

    Jacquot, Cyril; Szymanska, Jadwiga; Nemana, Lakshmi J; Steinbach, Lynne S; Horvai, Andrew E

    2015-11-01

    We present the case of a 46-year-old woman with no significant past medical history who developed left mid-thigh pain and fullness. Imaging demonstrated a mineralized soft-tissue mass, which increased in size during a year of monitoring, but retained a circumscribed appearance. The mass was located in the medial soft tissues of the thigh, separate from the bone on imaging studies, and this finding was confirmed during excision. The mass showed gross and microscopic features of an aneurysmal bone cyst. This diagnosis was supported by cytogenetic analysis revealing a t(17;17)(p13;q21) translocation corresponding to the USP6 and COL1A1 loci. Soft-tissue aneurysmal bone cyst is a rare entity, with fewer than 25 reports in the literature. Limited cytogenetic information about these tumors is available. To our knowledge, the USP6 and COL1A1 rearrangement has only previously been described in a pediatric soft-tissue aneurysmal bone cyst. We also discuss the differential diagnosis of ossifying soft-tissue lesions. PMID:26142538

  1. siRNA-mediated Allele-specific Silencing of a COL6A3 Mutation in a Cellular Model of Dominant Ullrich Muscular Dystrophy

    PubMed Central

    Bolduc, Véronique; Zou, Yaqun; Ko, Dayoung; Bönnemann, Carsten G

    2014-01-01

    Congenital muscular dystrophy type Ullrich (UCMD) is a severe disorder of early childhood onset for which currently there is no effective treatment. UCMD commonly is caused by dominant-negative mutations in the genes coding for collagen type VI, a major microfibrillar component of the extracellular matrix surrounding the muscle fibers. To explore RNA interference (RNAi) as a potential therapy for UCMD, we designed a series of small interfering RNA (siRNA) oligos that specifically target the most common mutations resulting in skipping of exon 16 in the COL6A3 gene and tested them in UCMD-derived dermal fibroblasts. Transcript analysis by semiquantitative and quantitative reverse transcriptase PCR showed that two of these siRNAs were the most allele-specific, i.e., they efficiently knocked down the expression from the mutant allele, without affecting the normal allele. In HEK293T cells, these siRNAs selectively suppressed protein expression from a reporter construct carrying the mutation, with no or minimal suppression of the wild-type (WT) construct, suggesting that collagen VI protein levels are as also reduced in an allele-specific manner. Furthermore, we found that treating UCMD fibroblasts with these siRNAs considerably improved the quantity and quality of the collagen VI matrix, as assessed by confocal microscopy. Our current study establishes RNAi as a promising molecular approach for treating dominant COL6-related dystrophies. PMID:24518369

  2. The heat-shock DnaK protein is required for plasmid R1 replication and it is dispensable for plasmid ColE1 replication.

    PubMed Central

    Giraldo-Suárez, R; Fernández-Tresguerres, E; Díaz-Orejas, R; Malki, A; Kohiyama, M

    1993-01-01

    Plasmid R1 replication in vitro is inactive in extracts prepared from a dnaK756 strain but is restored to normal levels upon addition of purified DnaK protein. Replication of R1 in extracts of a dnaKwt strain can be specifically inhibited with polyclonal antibodies against DnaK. RepA-dependent replication of R1 in dnaK756 extracts supplemented with DnaKwt protein at maximum concentration is partially inhibited by rifampicin and it is severely inhibited at sub-optimal concentrations of DnaK protein. The copy number of a run-away R1 vector is reduced in a dnaK756 background at 30 degrees C and at 42 degrees C the amplification of the run-away R1 vector is prevented. However a runaway R1 vector containing dnaK gene allows the amplification of the plasmid at high temperature. These data indicate that DnaK is required for both in vitro and in vivo replication of plasmid R1 and show a partial compensation for the low level of DnaK by RNA polymerase. In contrast ColE1 replication is not affected by DnaK as indicated by the fact that ColE1 replicates with the same efficiency in extracts from dnaKwt and dnaK756 strains. Images PMID:8265367

  3. Immobilization of cross linked Col-I-OPN bone matrix protein on aminolysed PCL surfaces enhances initial biocompatibility of human adipogenic mesenchymal stem cells (hADMSC)

    NASA Astrophysics Data System (ADS)

    Kim, Young-Hee; Jyoti, Md. Anirban; Song, Ho-Yeon

    2014-06-01

    In bone tissue engineering surface modification is considered as one of the important ways of fabricating successful biocompatible material. Addition of biologically active functionality on the surfaces has been tried for improving the overall biocompatibility of the system. In this study poly-ɛ-caprolactone film surfaces have been modified through aminolysis and immobilization process. Collagen type I (COL-I) and osteopontin (OPN), which play an important role in osteogenesis, was immobilized onto PCL films followed by aminolysis treatment using 1,6-hexanediamine. Characterization of animolysed and immobilized surfaces were done by a number techniques using scanning electron microscopy (SEM), FT-IR, XPS, ninhydrin staining, SDS-PAGE and confocal microscopy and compared between the modified and un-modified surfaces. Results of the successive experiments showed that aminolysis treatment was homogeneously achieved which helped to entrap or immobilize Col-I-OPN proteins on surfaces of PCL film. In vitro studies with human adipogenic mesenchymal stem cells (hADMSC) also confirmed the attachment and proliferation of cells was better in modified PCL surfaces than the unmodified surfaces. SEM, confocal microscopy and MTT assay showed a significant increase in cell spreading, attachment and proliferations on the biofunctionalized surfaces compared to the unmodified PCL surfaces at all-time points indicating the success of surface biofunctionalization.

  4. Molecular Genetic Diagnosis of a Bethlem Myopathy Family with an Autosomal-Dominant COL6A1 Mutation, as Evidenced by Exome Sequencing

    PubMed Central

    Park, Hyung Jun; Choi, Young-Chul; Kim, Seung Min; Kim, Se Hoon; Hong, Young Bin; Yoon, Bo Ram

    2015-01-01

    Background We describe herein the application of whole exome sequencing (WES) for the molecular genetic diagnosis of a large Korean family with dominantly inherited myopathy. Case Report The affected individuals presented with slowly progressive proximal weakness and ankle contracture. They were initially diagnosed with limb-girdle muscular dystrophy (LGMD) based on clinical and pathologic features. However, WES and subsequent capillary sequencing identified a pathogenic splicing-site mutation (c.1056+1G>A) in COL6A1, which was previously reported to be an underlying cause of Bethlem myopathy. After identification of the genetic cause of the disease, careful neurologic examination revealed subtle contracture of the interphalangeal joint in the affected members, which is a characteristic sign of Bethlem myopathy. Therefore, we revised the original diagnosis from LGMD to Bethlem myopathy. Conclusions This is the first report of identification of COL6A1-mediated Bethlem myopathy in Korea, and indicates the utility of WES for the diagnosis of muscular dystrophy. PMID:25749816

  5. Recessive and dominant mutations in COL12A1 cause a novel EDS/myopathy overlap syndrome in humans and mice

    PubMed Central

    Zou, Yaqun; Zwolanek, Daniela; Izu, Yayoi; Gandhy, Shreya; Schreiber, Gudrun; Brockmann, Knut; Devoto, Marcella; Tian, Zuozhen; Hu, Ying; Veit, Guido; Meier, Markus; Stetefeld, Jörg; Hicks, Debbie; Straub, Volker; Voermans, Nicol C.; Birk, David E.; Barton, Elisabeth R.; Koch, Manuel; Bönnemann, Carsten G.

    2014-01-01

    Collagen VI-related myopathies are disorders of connective tissue presenting with an overlap phenotype combining clinical involvement from the muscle and from the connective tissue. Not all patients displaying related overlap phenotypes between muscle and connective tissue have mutations in collagen VI. Here, we report a homozygous recessive loss of function mutation and a de novo dominant mutation in collagen XII (COL12A1) as underlying a novel overlap syndrome involving muscle and connective tissue. Two siblings homozygous for a loss of function mutation showed widespread joint hyperlaxity combined with weakness precluding independent ambulation, while the patient with the de novo missense mutation was more mildly affected, showing improvement including the acquisition of walking. A mouse model with inactivation of the Col12a1 gene showed decreased grip strength, a delay in fiber-type transition and a deficiency in passive force generation while the muscle seems more resistant to eccentric contraction induced force drop, indicating a role for a matrix-based passive force-transducing elastic element in the generation of the weakness. This new muscle connective tissue overlap syndrome expands on the emerging importance of the muscle extracellular matrix in the pathogenesis of muscle disease. PMID:24334604

  6. Enhanced Wnt signaling improves bone mass and strength, but not brittleness, in the Col1a1(+/mov13) mouse model of type I Osteogenesis Imperfecta.

    PubMed

    Jacobsen, Christina M; Schwartz, Marissa A; Roberts, Heather J; Lim, Kyung-Eun; Spevak, Lyudmila; Boskey, Adele L; Zurakowski, David; Robling, Alexander G; Warman, Matthew L

    2016-09-01

    Osteogenesis Imperfecta (OI) comprises a group of genetic skeletal fragility disorders. The mildest form of OI, Osteogenesis Imperfecta type I, is frequently caused by haploinsufficiency mutations in COL1A1, the gene encoding the α1(I) chain of type 1 collagen. Children with OI type I have a 95-fold higher fracture rate compared to unaffected children. Therapies for OI type I in the pediatric population are limited to anti-catabolic agents. In adults with osteoporosis, anabolic therapies that enhance Wnt signaling in bone improve bone mass, and ongoing clinical trials are determining if these therapies also reduce fracture risk. We performed a proof-of-principle experiment in mice to determine whether enhancing Wnt signaling in bone could benefit children with OI type I. We crossed a mouse model of OI type I (Col1a1(+/Mov13)) with a high bone mass (HBM) mouse (Lrp5(+/p.A214V)) that has increased bone strength from enhanced Wnt signaling. Offspring that inherited the OI and HBM alleles had higher bone mass and strength than mice that inherited the OI allele alone. However, OI+HBM and OI mice still had bones with lower ductility compared to wild-type mice. We conclude that enhancing Wnt signaling does not make OI bone normal, but does improve bone properties that could reduce fracture risk. Therefore, agents that enhance Wnt signaling are likely to benefit children and adults with OI type 1. PMID:27297606

  7. The arginine repressor is essential for plasmid-stabilizing site-specific recombination at the ColE1 cer locus.

    PubMed Central

    Stirling, C J; Szatmari, G; Stewart, G; Smith, M C; Sherratt, D J

    1988-01-01

    The heritable stability in Escherichia coli of the multicopy plasmid ColE1 and its natural relatives requires that the plasmids be maintained in the monomeric state. Plasmid multimers, that arise through recA-dependent homologous recombination, are normally converted to monomers by a site-specific recombination system that acts at a specific plasmid site (cer in ColE1). No plasmid functions that act at this site have been identified. In contrast, two unlinked E.coli genes that encode functions required for cer-mediated site-specific recombination have been identified. Here we describe the isolation and characterization of one such gene (xerA) and show it to be identical to the gene encoding the repressor of the arginine biosynthetic genes (argR). The argR protein binds to cer DNA both in vivo and in vitro in the presence of arginine. We believe this binding is required to generate a higher order protein-DNA complex within the recombinational synapse. The argR gene of Bacillus subtilis complements an E.coli argR deficiency for cer-mediated recombination despite the two proteins having only 27% amino acid identity. Images PMID:3149585

  8. Plasmids in the driving seat: The regulatory RNA Rcd gives plasmid ColE1 control over division and growth of its E. coli host.

    PubMed

    Gaimster, Hannah; Summers, David

    2015-03-01

    Regulation by non-coding RNAs was found to be widespread among plasmids and other mobile elements of bacteria well before its ubiquity in the eukaryotic world was suspected. As an increasing number of examples was characterised, a common mechanism began to emerge. Non-coding RNAs, such as CopA and Sok from plasmid R1, or RNAI from ColE1, exerted regulation by refolding the secondary structures of their target RNAs or modifying their translation. One regulatory RNA that seemed to swim against the tide was Rcd, encoded within the multimer resolution site of ColE1. Required for high fidelity maintenance of the plasmid in recombination-proficient hosts, Rcd was found to have a protein target, elevating indole production by stimulating tryptophanase. Rcd production is up-regulated in dimer-containing cells and the consequent increase in indole is part of the response to the rapid accumulation of dimers by over-replication (known as the dimer catastrophe). It is proposed that indole simultaneously inhibits cell division and plasmid replication, stopping the catastrophe and allowing time for the resolution of dimers to monomers. The idea of a plasmid-mediated cell division checkpoint, proposed but then discarded in the 1980s, appears to be enjoying a revival. PMID:25446541

  9. Stop codon in the procollagen II gene (COL2A1) in a family with the Stickler syndrome (arthro-ophthalmopathy).

    PubMed Central

    Ahmad, N N; Ala-Kokko, L; Knowlton, R G; Jimenez, S A; Weaver, E J; Maguire, J I; Tasman, W; Prockop, D J

    1991-01-01

    Linkage analysis with restriction fragment length polymorphisms for the gene for type II procollagen (COL2A1) was carried out in a family with the Stickler syndrome, or arthro-ophthalmopathy, an autosomal dominant disorder that affects the eyes, ears, joints, and skeleton. The analysis demonstrated linkage of the disease and COL2A1 with a logarithm-of-odds score of 1.51 at zero recombination. A newly developed procedure for preparing cosmid clones was employed to isolate the allele for type II procollagen that was linked to the disease. Analysis of over 7000 nucleotides of the gene revealed a single base mutation that altered a CG dinucleotide and converted the codon CGA for arginine at amino acid position alpha 1-732 to TGA, a stop codon. From previous work on procollagen biosynthesis, it is apparent that the truncated polypeptide synthesized from an allele with a stop codon at alpha 1-732 cannot participate in the assembly of type II procollagen, and therefore that the mutation would decrease synthesis of type II procollagen. It was not apparent, however, why the mutation produced marked changes in the eye, which contains only small amounts of type II collagen, but relatively mild effects on the many cartilaginous structures of the body that are rich in the same protein. Images PMID:1677770

  10. Mutations within the gene encoding the alpha 1 (X) chain of type X collagen (COL10A1) cause metaphyseal chondrodysplasia type Schmid but not several other forms of metaphyseal chondrodysplasia.

    PubMed Central

    Wallis, G A; Rash, B; Sykes, B; Bonaventure, J; Maroteaux, P; Zabel, B; Wynne-Davies, R; Grant, M E; Boot-Handford, R P

    1996-01-01

    Type X collagen is a homotrimer of alpha 1 (X) chains encoded by the COL10A1 gene. It is synthesised specifically and transiently by hypertrophic chondrocytes at sites of endochondral ossification. Point mutations and deletions in the region of the COL10A1 gene encoding the alpha 1 (X) carboxyl-terminal (NC1) domain have previously been identified in subjects with metaphyseal chondrodysplasia type Schmid (MCDS). To determine whether mutations in other regions of the gene caused MCDS or comparable phenotypes, we used PCR followed by SSCP to analyse the coding and promoter regions of the COL10A1 gene, as well as the intron/exon boundaries of five further subjects with MCDS, one subject with atypical MCDS, and nine subjects with other forms of metaphyseal chondrodysplasia. Using this approach, three of the subjects with MCDS were found to be heterozygous for the deletions 1864delACTT, 1956delT, and 2029delAC in the region of COL10A1 encoding the NC1 domain. These deletions would lead to alterations in the reading frame, premature stop codons, and the translation of truncated protein products. A fourth subject with MCDS was found to be heterozygous for a single base pair transition, T1894C, that would lead to the substitution of the amino acid residue serine at position 600 by proline within the NC1 domain. We did not, however, detect mutations in the coding and non-coding regions of COL10A1 in one subject with MCDS, the subject with atypical MCDS, and in the nine subjects with other forms of metaphyseal chondrodysplasia. We propose that the nature and distribution of mutations within the NC1 domain of COL10A1 causing MCDS argues against the hypothesis that the phenotype arises simply through haploinsufficiency but that an, as yet, unexplained mutation mechanism underlies this phenotype. Images PMID:8782043

  11. A rare haplotype in the upstream regulatory region of COL1A1 is associated with reduced bone quality and hip fracture.

    PubMed

    Jin, Huilin; Stewart, Tracy L; Hof, Rob Van't; Reid, David M; Aspden, Richard M; Ralston, Stuart

    2009-03-01

    Three polymorphisms have been identified in the 5' regulatory region of the COL1A1 gene at positions -1997 (rs1107946), -1663 (rs2412298), and +1245G/T (rs1800012), which combine to form haplotypes that have been associated with BMD in several populations. These polymorphisms and haplotypes have not thus far been studied in relation to biomechanical properties of bone or fracture risk. Genotypes and haplotypes of the COL1A1 gene were related to the biomechanical properties of bone ex vivo in samples of bone tissue obtained from the femoral head of 98 consecutive patients undergoing surgery for low-trauma hip fractures. Genotype and haplotype frequencies in the hip fracture cases were compared with 3418 population-based controls recruited from the same region. All three polymorphisms were associated with material density of the bone core, yield strength, and toughness. The association between -1663InsdelT and +1245G/T alleles, yield strength, and toughness remained significant after adjusting for material density of the core and other confounding factors. A haplotype comprising the unfavorable allele at all three polymorphic sites (-1997T/-1663delT/+1245T) was also associated with yield strength, modulus, and toughness after adjusting for confounding factors. This haplotype was carried by 19/94 (20.2%) patients with hip fracture compared with only 2/3399 (0.06%) female controls drawn from the general population (p < 0.0001). In contrast, there was no significant difference between cases and controls in genotype distribution for the individual polymorphisms. This study shows that common genetic variants in the 5' regulatory region of COL1A1 are associated with biomechanical properties of bone and reduced bone quality by mechanisms independent of their effects on BMD. The biomechanically unfavorable allele at each polymorphic site defines a haplotype that is extremely rare in the general population but that is approximately 400-fold enriched in hip fracture patients

  12. EndoS Reduces the Pathogenicity of Anti-mCOL7 IgG through Reduced Binding of Immune Complexes to Neutrophils

    PubMed Central

    Yu, Xinhua; Zheng, Junfeng; Collin, Mattias; Schmidt, Enno; Zillikens, Detlef; Petersen, Frank

    2014-01-01

    Endo-β-N-acetylglucosaminidase (EndoS) has been shown to act as a potent pathogen-derived immunomodulatory molecule in autoimmune diseases. Here we investigated how EndoS treatment reduces the pathogenicity of rabbit anti-mCOL7 IgG using different experimental models of epidermolysis bullosa acquisita (EBA). Our results show that the EndoS treatment does not interfere with the binding of the antibody to the antigen but reduces immune complex (IC)-mediated neutrophil activation by impairing the binding of the IC to FcγR on neutrophils. On the basis of this newly identified EndoS-mediated mechanism we hope to develop new strategies in the treatment of the disease. PMID:24504190

  13. Case Report: Whole exome sequencing reveals a novel frameshift deletion mutation p.G2254fs in COL7A1 associated with autosomal recessive dystrophic epidermolysis bullosa

    PubMed Central

    Karuthedath Vellarikkal, Shamsudheen; Jayarajan, Rijith; Verma, Ankit; Nair, Sreelata; Ravi, Rowmika; Senthivel, Vigneshwar; Sivasubbu, Sridhar; Scaria, Vinod

    2016-01-01

    Dystrophic epidermolysis bullosa simplex (DEB) is a phenotypically diverse inherited skin fragility disorder. It is majorly manifested by appearance of epidermal bullae upon friction caused either by physical or environmental trauma. The phenotypic manifestations also include appearance of milia, scarring all over the body and nail dystrophy. DEB can be inherited in a recessive or dominant form and the recessive form of DEB (RDEB) is more severe. In the present study, we identify a novel p.G2254fs mutation in COL7A1 gene causing a sporadic case of RDEB by whole exome sequencing (WES). Apart from adding a novel frameshift Collagen VII mutation to the repertoire of known mutations reported in the disease, to the best of our knowledge, this is the first report of a genetically characterized case of DEB from India. PMID:27408687

  14. Report of five novel and one recurrent COL2A1 mutations with analysis of genotype-phenotype correlation in patients with a lethal type II collagen disorder.

    PubMed

    Mortier, G R; Weis, M; Nuytinck, L; King, L M; Wilkin, D J; De Paepe, A; Lachman, R S; Rimoin, D L; Eyre, D R; Cohn, D H

    2000-04-01

    Achondrogenesis II-hypochondrogenesis and severe spondyloepiphyseal dysplasia congenita (SEDC) are lethal forms of dwarfism caused by dominant mutations in the type II collagen gene (COL2A1). To identify the underlying defect in seven cases with this group of conditions, we used the combined strategy of cartilage protein analysis and COL2A1 mutation analysis. Overmodified type II collagen and the presence of type I collagen was found in the cartilage matrix of all seven cases. Five patients were heterozygous for a nucleotide change that predicted a glycine substitution in the triple helical domain (G313S, G517V, G571A, G910C, G943S). In all five cases, analysis of cartilage type II collagen suggested incorporation of the abnormal alpha1(II) chain in the extracellular collagen trimers. The G943S mutation has been reported previously in another unrelated patient with a strikingly similar phenotype, illustrating the possible specific effect of the mutation. The radiographically less severely affected patient was heterozygous for a 4 bp deletion in the splice donor site of intron 35, likely to result in aberrant splicing. One case was shown to be heterozygous for a single nucleotide change predicted to result in a T1191N substitution in the carboxy-propeptide of the proalpha1(II) collagen chain. Study of the clinical, radiographic, and morphological features of the seven cases supports evidence for a phenotypic continuum between achondrogenesis II-hypochondrogenesis and lethal SEDC and suggests a relationship between the amount of type I collagen in the cartilage and the severity of the phenotype. PMID:10745044

  15. An RNA-splicing mutation (G+5IVS20) in the type II collagen gene (COL2A1) in a family with spondyloepiphyseal dysplasia congenita.

    PubMed

    Tiller, G E; Weis, M A; Polumbo, P A; Gruber, H E; Rimoin, D L; Cohn, D H; Eyre, D R

    1995-02-01

    Defects in type II collagen have been demonstrated in a phenotypic continuum of chondrodysplasias that includes achondrogenesis II, hypochondrogenesis, spondyloepiphyseal dysplasia congenita (SEDC), Kniest dysplasia, and Stickler syndrome. We have determined that cartilage from a terminated fetus with an inherited form of SEDC contained both normal alpha 1(II) collagen chains and chains that lacked amino acids 256-273 of the triple-helical domain. PCR amplification of this region of COL2A1, from genomic DNA, yielded products of normal size, while amplification of cDNA yielded a normal sized species and a shorter fragment missing exon 20. Sequence analysis of genomic DNA from the fetus revealed a G-->T transversion at position +5 of intron 20; the affected father was also heterozygous for the mutation. Allele-specific PCR and heteroduplex analysis of a VNTR in COL2A1 independently confirmed the unaffected status of a fetus in a subsequent pregnancy. Thermodynamic calculations suggest that the mutation prevents normal splicing of exon 20 by interfering with binding of U1 small-nuclear RNA to pre-mRNA, thus leading to skipping of exon 20 in transcripts from the mutant allele. Electron micrographs of diseased cartilage showed intracellular inclusion bodies, which were stained by an antibody to alpha 1(II) procollagen. Our findings support the hypothesis that alpha-chain length alterations that preserve the Gly-X-Y repeat motif of the triple helix result in partial intracellular retention of alpha 1(II) procollagen and produce mild to moderate chondrodysplasia phenotypes. PMID:7847372

  16. An RNA-splicing mutation (G{sup +51VS20}) in the Type II collagen gene (COL2A1) in a family with spondyloepiphyseal dysplasia congenita

    SciTech Connect

    Tiller, G.E.; Polumbo, P.A.; Weis, M.A.; Eyre, D.R.; Gruber, H.E.; Rimoin, D.L.; Cohn, D.H. |

    1995-02-01

    Defects in type II collagen have been demonstrated in a phenotypic continuum of chondrodysplasias that includes achondrogenesis II, hypochondrogenesis, spondyloepiphyseal dysplasia congenita (SEDC), Kniest dysplasia, and Stickler syndrome. We have determined that cartilage from a terminated fetus with an inherited form of SEDC contained both normal {alpha}1(II) collagen chains and chains that lacked amino acids 256-273 of the triple-helical domain. PCR amplification of this region of COL2A1, from genomic DNA, yielded products of normal size, while amplification of cDNA yielded a normal sized species and a shorter fragment missing exon 20. Sequence analysis of genomic DNA from the fetus revealed a G{yields}T transversion at position +5 of intron 20; the affected father was also heterozygous for the mutation. Allele-specific PCR and heteroduplex analysis of a VNTR in COL2A1 independently confirmed the unaffected status of a fetus in a subsequent pregnancy. Thermodynamic calculations suggest that the mutation prevents normal splicing of exon 20 by interfering with binding of U{sub 1} small-nuclear RNA to pre-mRNA, thus leading to skipping of exon 20 in transcripts from the mutant allele. Electron micrographs of diseased cartilage showed intracellular inclusion bodies, which were stained by an antibody to {alpha}1(II) procollagen. Our findings support the hypothesis that {alpha}-chain length alterations that preserve the Gly-X-Y repeat motif of the triple helix result in partial intracellular retention of {alpha}1(II) procollagen and produce mild to moderate chondrodysplasia phenotypes. 50 refs., 6 figs., 1 tab.

  17. A 1750-1999 AD Atmospheric Record of East Asian Black Carbon Emissions from the Prospector-Russell Col Ice Core, Mt. Logan, Yukon Territory

    NASA Astrophysics Data System (ADS)

    Menking, J. A.; Kaspari, S.; Osterberg, E. C.; Fisher, D.

    2012-12-01

    Black carbon (BC), an aerosol created from the incomplete combustion of fossil and biofuels, absorbs sunlight causing warming in the atmosphere and cooling at the surface. BC deposited on snow and ice reduces the surface albedo, accelerating seasonal snowmelt and glacier retreat and influencing the regional water cycle and climate. Estimates place East Asia as the top BC producer worldwide. Recent studies suggest that increases in East Asian BC related to industrialization have influenced Asian climate by altering temperatures and the timing and magnitude of precipitation. East Asian BC emissions have been estimated using fossil fuel inventories, but observational data is needed to constrain the timing and magnitude of recent BC emissions changes in order to better assess the role of BC in recent climate change. Here we present a time series of East Asian atmospheric BC that spans 1750-1999 AD that was developed using an ice core collected at the Prospector-Russell (PR) Col on Mt. Logan, Yukon Territory, Canada. The PR Col ice core is ideal for developing a record of East Asian BC emissions because the Mt. Logan summit has been shown to capture Asian emissions transported across the Pacific Ocean. The Mt. Logan record shows increases in BC concentration occurring in the early 1980's related to the recent industrialization of East Asian nations. We utilize the BC record from Mt. Logan (1) for comparison with estimated East Asian emissions inventories, and (2) to assess the role of BC in recent climate change with respect to the timing and magnitude of changes in East Asian BC emissions.

  18. SEARCHING FOR YOUNG JUPITER ANALOGS AROUND AP COL: L-BAND HIGH-CONTRAST IMAGING OF THE CLOSEST PRE-MAIN-SEQUENCE STAR

    SciTech Connect

    Quanz, Sascha P.; Avenhaus, Henning; Meyer, Michael R.; Crepp, Justin R.; Hillenbrand, Lynne A.; Janson, Markus

    2012-08-01

    The nearby M-dwarf AP Col was recently identified by Riedel et al. as a pre-main-sequence star (age 12-50 Myr) situated only 8.4 pc from the Sun. The combination of its youth, distance, and intrinsically low luminosity make it an ideal target to search for extrasolar planets using direct imaging. We report deep adaptive optics observations of AP Col taken with VLT/NACO and Keck/NIRC2 in the L band. Using aggressive speckle suppression and background subtraction techniques, we are able to rule out companions with mass m {>=} 0.5-1 M{sub Jup} for projected separations a > 4.5 AU, and m {>=} 2 M{sub Jup} for projected separations as small as 3 AU, assuming an age of 40 Myr using the COND theoretical evolutionary models. Using a different set of models, the mass limits increase by a factor of {approx}>2. The observations presented here are the deepest mass-sensitivity limits yet achieved within 20 AU on a star with direct imaging. While Doppler radial velocity surveys have shown that Jovian bodies with close-in orbits are rare around M-dwarfs, gravitational microlensing studies predict that 17{sup +6}{sub -9}% of these stars host massive planets with orbital separations of 1-10 AU. Sensitive high-contrast imaging observations, like those presented here, will help to validate results from complementary detection techniques by determining the frequency of gas giant planets on wide orbits around M-dwarfs.

  19. The type of variants at the COL3A1 gene associates with the phenotype and severity of vascular Ehlers-Danlos syndrome.

    PubMed

    Frank, Michael; Albuisson, Juliette; Ranque, Brigitte; Golmard, Lisa; Mazzella, Jean-Michael; Bal-Theoleyre, Laurence; Fauret, Anne-Laure; Mirault, Tristan; Denarié, Nicolas; Mousseaux, Elie; Boutouyrie, Pierre; Fiessinger, Jean-Noël; Emmerich, Joseph; Messas, Emmanuel; Jeunemaitre, Xavier

    2015-12-01

    Vascular Ehlers-Danlos syndrome (vEDS) is a rare and severe autosomal dominant disorder caused by variants at the COL3A1 gene. Clinical characteristics and course of disease of 215 molecularly proven patients (146 index cases and 69 relatives) were analysed. We found 126 distincts variants that were divided into five groups: (1) Glycine substitutions (n=71), (2) splice-site and in-frame insertions-deletions (n=36), (3) variants leading to haplo-insufficiency (n=7), (4) non-glycine missense variants within the triple helix (n=4 variants), and (5) non-glycine missense variants or in-frame insertions-deletions, in the N- or C-terminal part of the protein (n=8). Overall, our cohort confirmed the severity of the disease with a median age at first complication of 29 years (IQR 22-39), the most frequent being arterial (48%) and digestive (24%) ruptures. Groups 2 and 1 were significantly more severe than groups 3-5, with extreme median ages at first major complication of 23-47 years. Patients of groups 3-5 had a less typical phenotype and remarkably absence of digestive events. The distribution of glycine-replacing amino acids was strongly biased towards more destabilizing residues of the collagen assembly. Thus the natural course of vEDS and the clinical phenotype of patients are influenced by the type of COL3A1 variant. This study also confirms that patients with variants located in the C- and N-termini or leading to haplo-insufficiency have milder course of the disease and less prevalent diagnostic criteria. These findings may help refine diagnostic strategy, genetic counselling and clinical care. PMID:25758994

  20. miR-29a/b enhances cell migration and invasion in nasopharyngeal carcinoma progression by regulating SPARC and COL3A1 gene expression.

    PubMed

    Qiu, Feifei; Sun, Rui; Deng, Ning; Guo, Tianyu; Cao, Yange; Yu, Ying; Wang, Xuejun; Zou, Bingcheng; Zhang, Songmei; Jing, Tao; Ling, Tao; Xie, Jun; Zhang, Qing

    2015-01-01

    Nasopharyngeal carcinoma (NPC) is a malignant tumor associated with a genetic predisposition, Epstein-Barr virus infection and chromosomal abnormalities. Recently, several miRNAs have been shown to target specific mRNAs to regulate NPC development and progression. However, the involvement of miRNAs in processes leading to NPC migration and invasion remains to be elucidated. We predicted that miR-29a/b are associated with dysregulated genes controlling NPC through an integrated interaction network of miRNAs and genes. miR-29a/b over-expression in NPC cell lines had no significant effect on proliferation, whereas miR-29b mildly increased the percentage of cells in the G1 phase with a concomitant decrease in the percentage of cells in S phase. Furthermore, we demonstrated that miR-29a/b might be responsible for increasing S18 cell migration and invasion, and only COL3A1 was identified as a direct target of miR-29b despite the fact that both SPARC and COL3A1 were inhibited by miR-29a/b over-expression. Meanwhile, SPARC proteins were increased in metastatic NPC tissue and are involved in NPC progression. Unexpectedly, we identified that miRNA-29b expression was elevated in the serum of NPC patients with a high risk of metastasis. The 5-year actuarial overall survival rates in NPC patients with high serum miR-29b expression was significantly shorter than those with low serum miR-29b expression; therefore, serum miR-29b expression could be a promising prognostic marker. PMID:25786138

  1. Association of polymorphism at COL3A and CTLA4 loci on chromosome 2q31-33 with the clinical phenotype and in-vitro CMI status in healthy and leprosy subjects: a preliminary study.

    PubMed

    Kaur, G; Sachdeva, G; Bhutani, L K; Bamezai, R

    1997-07-01

    Two genetic loci, viz. COL3A and CTLA4, located within the chromosome 2q31-33 region in the vicinity of the proposed syntenic site of the mouse "Bcg" locus were genotyped by the polymerase chain reaction in leprosy patients and healthy individuals. All the subjects studied were assessed as in-vitro responders/non-responders to mycobacterial antigens. Simple sequence length polymorphism analysis revealed five (236 to 312 bp) and eight (84 to 120 bp) allelomorphs for COL3A and CTLA4, respectively. Our preliminary analysis showed a significant association between the 250-bp COL3A allelomorph in the homozygous condition and the multibacillary form of leprosy (P < 0.05: relative risk = 5.5). Another allelic (312 bp) variant of COL3A was significantly correlated with non-responsiveness to M. leprae antigens in vitro (P < 0.01). The 104-bp allelomorph of CTLA4 was not observed in any of the 25 cases of leprosy. This absence was statistically significant (P < 0.05) when compared with normal healthy controls and depicted a high relative risk (RR = 25.83). An additional observation of the predominance of a unique 84-bp CTLA4/CTLA4-like allelomorph was observed in the Indian subjects studied. PMID:9225967

  2. A highly polymorphic (ACT)n VNTR (variable nucleotide of tandem repeats) locus inside intron 12 of COL1A2, one of the two genes involved in dominant osteogenesis imperfecta.

    PubMed

    Pepe, G

    1993-01-01

    A new, highly polymorphic, region consisting of variable number of tandem repeats (VNTR) is described that occurs within intron 12 of the COL1A2 gene. This VNTR consists of the trinucleotide ACT repeated from 6 to 12 times. Of the six alleles so far detected four are common in the three major races. The two rare alleles, (ACT)11 and (ACT)12, have been found only in Africans. In addition, a rapid technique has been developed that can be used successfully with very small amounts of even partially degraded DNA, thus allowing the use of this VNTR for forensic applications. Since dominant OI can be due to mutations at either of two loci (COL1A1 and COL1A2) prenatal diagnosis becomes feasible in the majority of the affected families only if a very informative marker is available for both of these genes. This VNTR provides a very powerful marker for COL1A2. In fact the heterozygosity for it ranges from 0.634 to 0.741 with PIC values from 0.562 to 0.696, respectively. Since trinucleotide repeats can be "unstable," and sometimes pathogenic, the unexplained collagenopathies (or suspected collagenopathies) should be analyzed from this point of view. PMID:8104634

  3. Whole Exome Sequencing and Segregation Analysis Confirms That a Mutation in COL17A1 Is the Cause of Epithelial Recurrent Erosion Dystrophy in a Large Dominant Pedigree Previously Mapped to Chromosome 10q23-q24

    PubMed Central

    Le, Derek J.; Chen, Yabin; Wang, Qiwei; Chung, D. Doug; Frausto, Ricardo F.; Croasdale, Christopher; Yee, Richard W.; Hejtmancik, Fielding J.; Aldave, Anthony J.

    2016-01-01

    Purpose To report identification of a COL17A1 mutation in a family with a corneal dystrophy previously mapped to chromosome 10q23-q24. Methods Whole-exome sequencing was performed on DNA samples from five affected family members and two unrelated, unaffected individuals. Identified variants were filtered for those that were: located in the linked interval on chromosome 10q23-q24; novel or rare (minor allele frequency ≤0.01); heterozygous; present in all affected individuals and not in controls; and present in genes that encode proteins expressed in human corneal epithelial cells (reads per kilobase per million ≥1). Sanger sequencing of identified variants (SNVs) was performed in additional family members. In silico analysis was used to predict the functional impact of non-synonymous variants. Results Three SNVs located in two genes were identified that met the filtering criteria: one rare synonymous c.3156C>T variant in the collagen, type XVII, alpha I (COL17A1) gene; and two rare variants, one synonymous and one missense, in the dynamin binding protein (DNMBP) gene. Sanger sequencing of additional family members determined that only the COL17A1 variant segregates with the affected phenotype. In silico analysis predicts that the missense variant in DNMBP would be tolerated. Conclusions The corneal dystrophy mapped to chromosome 10q23-q24 is associated with the c.3156C>T variant in COL17A1. As this variant has recently been identified in five other families with early onset recurrent corneal erosions, and has been shown in vitro to introduce a cryptic splice donor site, this dystrophy is likely caused by aberrant splicing of COL17A1 and should be classified as epithelial recurrent erosion dystrophy. PMID:27309958

  4. Using Visual Interpretive Analysis in Teaching Joyce Carol Oates's "Naked."

    ERIC Educational Resources Information Center

    Brandt, Kenneth K.

    2003-01-01

    Shares some of the classroom practices that have worked well in teaching "Naked." Presents a summary of introductory comments about the story and a description of the sketching activity. Offers interpretive commentary on a selection of representative student sketches that reflect significant features of the story. Provides general comments to…

  5. A Reflective Conversation with Joyce Van Tassel Baska

    ERIC Educational Resources Information Center

    Shaughnessy, Michael F.; Jager, Kaarina

    2012-01-01

    In an increasingly multicultural world, with increasing access to other countries and opportunities for foreign travel, gifted students need to be able to communicate with other students and other individuals around the world. If they engage in business endeavors, gifted entrepreneurs need to be able to communicate both verbally and in writing…

  6. Les facteurs pronostiques de survie sans récidive chez les patientes atteintes de tumeur du col de l'utérus

    PubMed Central

    Dossou, Serpos; James, Laurianne; Bakkali, Hanae; Afif, Mohammed; Rahali, Leila; Irigo, Joelle; Ogandaga, Etienne; Kebdani, Tayeb; Ahid, Samir; Benjaafar, Noureddine

    2015-01-01

    La radiochimiothérapie est le traitement de référence des tumeurs du col localement avancées, et plusieurs études ont montré l'importance des facteurs pronostiques sur le contrôle local de la tumeur et la survie des malades. L'objectif de cette étude est d'évaluer l'impact des facteurs pronostiques, en particulier de l'étalement sur la survie sans récidive des patientes suivies pour cancer du col utérin. Il s'agit d'une série rétrospective portant sur 177 femmes suivies en 2011 pour tumeur du col utérin de stade IB à III selon la classification de FIGO 2009 ayant bénéficié d'une radiothérapie à la dose de 46 Grays sur le pelvis et une surimpression sur les paramètres envahis associée à du cisplatine 40mg/m2 par semaine suivie de curiethérapie réalisée selon le mode haut débit de dose (HDR) ou bas débit de dose (LDR). La moyenne d'âge était de 53ans, la médiane de l'étalement total était de 65 jours, 75% des patientes ont reçu 4 cures de chimiothérapie, et les patientes ont été suivies après le traitement pendant une durée médiane de 34 mois. La récidive locale et métastatique était de 33,3% chez les patientes ayant des adénopathies pelviennes, contre 16,3% chez celles qui en étaient indemnes (p= 0,031), elle était de 26,3% chez les patientes ayant un étalement supérieur à 65 jours contre 11% chez celles dont l'étalement en était inférieur (p= 0,01). La présence d'adénopathies pelviennes et l'étalement total de la radiothérapie apparaissaient respectivement comme les seuls facteurs pronostiques indépendant de survenue de récidive, p= 0,04 OR= 2,6 IC95% (1,05 6,3) et p= 0,01 OR= 2,9 IC95% (1,26 6,7). En analyse multivariée, la technique de curiethérapie p = 0,003 OR= 0,25 IC95% (0,1 0,6) et l'étalement total du traitement p= 0,0001 OR= 4,7 IC95% (2 10,8) apparaissaient comme les seuls facteurs pronostiques indépendant de survie sans récidive. L'étalement supérieur à 65 jours et la technique de curieth

  7. Improving the Terrain-Based Parameter for the Assessment of Snow Redistribution in the Col du Lac Blanc Area and Comparisons with TLS Snow Depth Data

    NASA Astrophysics Data System (ADS)

    Schön, Peter; Prokop, Alexander; Naaim-Bouvet, Florence; Nishimura, Kouichi; Vionnet, Vincent; Guyomarc'h, Gilbert

    2014-05-01

    Wind and the associated snow drift are dominating factors determining the snow distribution and accumulation in alpine areas, resulting in a high spatial variability of snow depth that is difficult to evaluate and quantify. The terrain-based parameter Sx characterizes the degree of shelter or exposure of a grid point provided by the upwind terrain, without the computational complexity of numerical wind field models. The parameter has shown to qualitatively predict snow redistribution with good reproduction of spatial patterns, but has failed to quantitatively describe the snow redistribution, and correlations with measured snow heights were poor. The objective of our research was to a) identify the sources of poor correlations between predicted and measured snow re-distribution and b) improve the parameters ability to qualitatively and quantitatively describe snow redistribution in our research area, the Col du Lac Blanc in the French Alps. The area is at an elevation of 2700 m and particularly suited for our study due to its constant wind direction and the availability of data from a meteorological station. Our work focused on areas with terrain edges of approximately 10 m height, and we worked with 1-2 m resolution digital terrain and snow surface data. We first compared the results of the terrain-based parameter calculations to measured snow-depths, obtained by high-accuracy terrestrial laser scan measurements. The results were similar to previous studies: The parameter was able to reproduce observed patterns in snow distribution, but regression analyses showed poor correlations between terrain-based parameter and measured snow-depths. We demonstrate how the correlations between measured and calculated snow heights improve if the parameter is calculated based on a snow surface model instead of a digital terrain model. We show how changing the parameter's search distance and how raster re-sampling and raster smoothing improve the results. To improve the parameter

  8. Three New Genetic Loci (R1210C in CFH, Variants in COL8A1 and RAD51B) Are Independently Related to Progression to Advanced Macular Degeneration

    PubMed Central

    Seddon, Johanna M.; Reynolds, Robyn; Yu, Yi; Rosner, Bernard

    2014-01-01

    Objectives To assess the independent impact of new genetic variants on conversion to advanced stages of AMD, controlling for established risk factors, and to determine the contribution of genes in predictive models. Methods In this prospective longitudinal study of 2765 individuals, 777 subjects progressed to neovascular disease (NV) or geographic atrophy (GA) in either eye over 12 years. Recently reported genetic loci were assessed for their independent effects on incident advanced AMD after controlling for 6 established loci in 5 genes, and demographic, behavioral, and macular characteristics. New variants which remained significantly related to progression were then added to a final multivariate model to assess their independent effects. The contribution of genes to risk models was assessed using reclassification tables by determining risk within cross-classified quintiles for alternative models. Results Three new genetic variants were significantly related to progression: rare variant R1210C in CFH (hazard ratio (HR) 2.5, 95% confidence interval [CI] 1.2–5.3, P = 0.01), and common variants in genes COL8A1 (HR 2.0, 95% CI 1.1–3.5, P = 0.02) and RAD51B (HR 0.8, 95% CI 0.60–0.97, P = 0.03). The area under the curve statistic (AUC) was significantly higher for the 9 gene model (.884) vs the 0 gene model (.873), P = .01. AUC’s for the 9 vs 6 gene models were not significantly different, but reclassification analyses indicated significant added information for more genes, with adjusted odds ratios (OR) for progression within 5 years per one quintile increase in risk score of 2.7, P<0.001 for the 9 vs 6 loci model, and OR 3.5, P<0.001 for the 9 vs. 0 gene model. Similar results were seen for NV and GA. Conclusions Rare variant CFH R1210C and common variants in COL8A1 and RAD51B plus six genes in previous models contribute additional predictive information for advanced AMD beyond macular and behavioral phenotypes. PMID:24498017

  9. Demise of the northern Tethyan Urgonian carbonate platform and subsequent transition towards pelagic conditions: The sedimentary record of the Col de la Plaine Morte area, central Switzerland

    NASA Astrophysics Data System (ADS)

    Föllmi, Karl B.; Gainon, François

    2008-04-01

    The sedimentary succession of the Col de la Plaine Morte area (Helvetic Alps, central Switzerland) documents the disappearance of the northern Tethyan Urgonian platform in unprecedented detail and suggests stepwise platform demise, with each drowning phase documented by erosion and phosphogenesis. The first identified drowning phase terminated Urgonian carbonate production in a predominantly photozoan mode. Using a correlation of the whole-rock δ13C record with the well-dated record from SE France, its age is inferred to as Middle Early Aptian (near the boundary between the weissi and deshayesi zones). A subsequent drowning phase is dated by ammonites and by a correlation of the whole-rock δ13C record as Late Early Aptian (late deshayesi to early furcata zone). A third drowning phase provides an ammonite-based age of Early Late Aptian ( subnodosocostatum and melchioris zones) and is part of a widely recognized phase of sediment condensation and phosphogenesis, which is dated as latest Early to Middle Late Aptian (late furcata zone to near the boundary of the melchioris and nolani zones). The fourth and final drowning phase started in the latest Aptian ( jacobi zone) as is also indicated by ammonite findings at the Col de la Plaine Morte. The phases of renewed platform-carbonate production intervening between the drowning phases were all in a heterozoan mode. During the ultimate drowning phase, phosphogenesis continued until the Early Middle Albian, whereas condensation processes lasted until the Middle Turonian. Coverage of the external margin of the drowned Urgonian platform by a drape of pelagic carbonates started only in the Late Turonian. During the Santonian, the external part of the drowned platform underwent normal faulting and saw the re-exposure of already lithified Urgonian carbonates at the seafloor. Based on the here-inferred ages, the first drowning phase just precedes oceanic anoxic episode 1a (OAE 1a or "selli event") in time, and the second

  10. Molecular Mining of Alleles in Water Buffalo Bubalus bubalis and Characterization of the TSPY1 and COL6A1 Genes

    PubMed Central

    Kumar, Sudhir; Ali, Sher

    2011-01-01

    Background Minisatellites are an integral part of eukaryotic genomes and show variation in the complexity of their organization. Besides their presence in non-coding regions, a small fraction of them are part of the transcriptome, possibly participating in gene regulation, expression and silencing. We studied the minisatellite (TGG)n tagged transcriptome in the water buffalo Bubalus bubalis across various tissues and the spermatozoa, and characterized the genes TSPY1 and COL6A1 discovered in the process. Results Minisatellite associated sequence amplification (MASA) conducted using cDNA and oligonucleotide primer (TGG)5 uncovered 38 different mRNA transcripts from somatic tissues and gonads and 15 from spermatozoa. These mRNA transcripts corresponded to several known and novel genes. The majority of the transcripts showed the highest level of expression either in the testes or spermatozoa with exception of a few showing higher expression levels in the lungs and liver. Transcript SR1, which is expressed in all the somatic tissues and gonads, was found to be similar to the Bos taurus collagen type VI alpha 1 gene (COL6A1). Similarly, SR29, a testis-specific transcript, was found to be similar to the Bos taurus testis-specific Y-encoded protein-1 representing cancer/testis antigen 78 (CT78). Subsequently, full length coding sequences (cds) of these two transcripts were obtained. Quantitative PCR (q-PCR) revealed 182-202 copies of theTSPY1 gene in water buffalo, which localized to the Y chromosome. Conclusions The MASA approach enabled us to identify several genes, including two of clinical significance, without screening an entire cDNA library. Genes identified with TGG repeats are not part of a specific family of proteins and instead are distributed randomly throughout the genome. Genes showing elevated expression in the testes and spermatozoa may prove to be potential candidates for in-depth characterization. Furthermore, their possible involvement in fertility or

  11. Merging a Terrain-Based Parameter and Snow Particle Counter Data for the Assessment of Snow Redistribution in the Col du Lac Blanc Area

    NASA Astrophysics Data System (ADS)

    Schön, Peter; Prokop, Alexander; Naaim-Bouvet, Florence; Vionnet, Vincent; Guyomarc'h, Gilbert; Heiser, Micha; Nishimura, Kouichi

    2015-04-01

    Wind and the associated snow drift are dominating factors determining the snow distribution and accumulation in alpine areas, resulting in a high spatial variability of snow depth that is difficult to evaluate and quantify. The terrain-based parameter Sx characterizes the degree of shelter or exposure of a grid point provided by the upwind terrain, without the computational complexity of numerical wind field models. The parameter has shown to qualitatively predict snow redistribution with good reproduction of spatial patterns. It does not, however, provide a quantitative estimate of changes in snow depths. The objective of our research was to introduce a new parameter to quantify changes in snow depths in our research area, the Col du Lac Blanc in the French Alps. The area is at an elevation of 2700 m and particularly suited for our study due to its consistently bi-modal wind directions. Our work focused on two pronounced, approximately 10 m high terrain breaks, and we worked with 1 m resolution digital snow surface models (DSM). The DSM and measured changes in snow depths were obtained with high-accuracy terrestrial laser scan (TLS) measurements. First we calculated the terrain-based parameter Sx on a digital snow surface model and correlated Sx with measured changes in snow-depths (Δ SH). Results showed that Δ SH can be approximated by Δ SHestimated = α * Sx, where α is a newly introduced parameter. The parameter α has shown to be linked to the amount of snow deposited influenced by blowing snow flux. At the Col du Lac Blanc test side, blowing snow flux is recorded with snow particle counters (SPC). Snow flux is the number of drifting snow particles per time and area. Hence, the SPC provide data about the duration and intensity of drifting snow events, two important factors not accounted for by the terrain parameter Sx. We analyse how the SPC snow flux data can be used to estimate the magnitude of the new variable parameter α . To simulate the development

  12. A COL2A1 mutation in achondrogenesis type II results in the replacement of type II collagen by type I and III collagens in cartilage.

    PubMed

    Chan, D; Cole, W G; Chow, C W; Mundlos, S; Bateman, J F

    1995-01-27

    An autosomal dominant mutation in the COL2A1 gene was identified in a fetus with achondrogenesis type II. A transition of G2853 to A in exon 41 produced a substitution of Gly769 by Ser within the triple helical domain of the alpha 1(II) chain of type II collagen, interrupting the mandatory Gly-X-Y triplet sequence required for the normal formation of stable triple helical type II collagen molecules, resulting in the complete absence of type II collagen in the cartilage, which had a gelatinous composition. Type I and III collagens were the major species found in cartilage tissue and synthesized by cultured chondrocytes along with cartilage type XI collagen. However, cultured chondrocytes produced a trace amount of type II collagen, which was retained within the cells and not secreted. In situ hybridization of cartilage sections showed that the chondrocytes produced both type II and type I collagen mRNA. As a result, it is likely that the chondrocytes produced type II collagen molecules, which were then degraded. The close proximity of the Gly769 substitution by Ser to the mammalian collagenase cleavage site at Gly775-Leu776 may have produced an unstable domain that was highly susceptible to proteolysis. The type I and III collagens that replaced type II collagen were unable to maintain the normal structure of the hyaline cartilage but did support chondrocyte maturation, evidenced by the expression of type X collagen in the hypertrophic zone of the growth plate cartilage. PMID:7829510

  13. Defective splicing of mRNA from one COL1A1 allele of type I collagen in nondeforming (type I) osteogenesis imperfecta.

    PubMed Central

    Stover, M L; Primorac, D; Liu, S C; McKinstry, M B; Rowe, D W

    1993-01-01

    Osteogenesis imperfecta (OI) type I is the mildest form of heritable bone fragility resulting from mutations within the COL1A1 gene. We studied fibroblasts established from a child with OI type I and demonstrated underproduction of alpha 1 (I) collagen chains and alpha 1 (I) mRNA. Indirect RNase protection suggested two species of alpha 1 (I) mRNA, one of which was not collinear with fully spliced alpha 1 (I) mRNA. The noncollinear population was confined to the nuclear compartment of the cell, and contained the entire sequence of intron 26 and a G-->A transition in the first position of the intron donor site. The G-->A transition was also identified in the genomic DNA. The retained intron contained an in-frame stop codon and introduced an out-of-frame insertion within the collagen mRNA producing stop codons downstream of the insertion. These changes probably account for the failure of the mutant RNA to appear in the cytoplasm. Unlike other splice site mutations within collagen mRNA that resulted in exon skipping and a truncated but inframe RNA transcript, this mutation did not result in production of a defective collagen pro alpha 1 (I) chain. Instead, the mild nature of the disease in this case reflects failure to process the defective mRNA and thus the absence of a protein product from the mutant allele. Images PMID:8408653

  14. Mutations in the 180-kD bullous pemphigoid antigen (BPAG2), a hemidesmosomal transmembrane collagen (COL17A1), in generalized atrophic benign epidermolysis bullosa.

    PubMed

    McGrath, J A; Gatalica, B; Christiano, A M; Li, K; Owaribe, K; McMillan, J R; Eady, R A; Uitto, J

    1995-09-01

    Junctional epidermolysis bullosa (JEB) is a heterogeneous autosomal recessively inherited blistering skin disorder associated with fragility at the dermal-epidermal junction. Characteristic ultrastructural findings in JEB are abnormalities in the hemidesmosome-anchoring filament complexes. These focal attachment structures, which extend from the intracellular compartment of the basal keratinocytes to the underlying basement membrane, have been shown to be hypoplastic or rudimentary in different forms of JEB. Previously, in different JEB phenotypes, mutations have been found in the three genes for the anchoring filament component laminin 5 (LAMA3, LAMB3, and LAMC2) and in the gene for the hemidesmosome-associated integrin beta 4 subunit. Here, we describe the first mutations in the gene encoding the 180-kD bullous pemphigoid antigen (BPAG2), a transmembranous hemidesmosomal collagen, also known as type XVII collagen (COL17A1). The patient is affected with generalized atrophic benign epidermolysis bullosa (GABEB), a rare variant of JEB, and is a compound heterozygote for premature termination codons on both alleles. These novel findings emphasize the molecular heterogeneity of this group of genodermatoses, and attest to the importance of BPAG2 in maintaining adhesion between the epidermis and the dermis. PMID:7550320

  15. Partial isodisomy for maternal chromosome 7 and short stature in an individual with a mutation at the COL1A2 locus

    SciTech Connect

    Spotila, L.D.; Sereda, L.; Prockop, D.J. )

    1992-12-01

    Uniparental disomy for chromosome 7 has been described previously in two individuals with cystic fibrosis. Here, the authors describe a third case that was discovered because the proband was homozygous for a mutation in the COL1A2 gene for type I procollagen, although his mother was heterozygous and his father did not have the mutation. Phenotypically, the proband was similar to the two previously reported cases with uniparental disomy for chromosome 7, in that he was short in stature and growth retarded. Paternity was assessed with five polymorphic markers. Chromosome 7 inheritance in the proband was analyzed using 12 polymorphic markers distributed along the entire chromosome. Similar analysis of the proband's two brothers established the phase of the alleles at the various loci, assuming minimal recombination. The proband inherited only maternal alleles at five loci and was homozygous at all loci examined, except one. He was heterozygous for an RFLP at the IGBP-1 locus at 7p13-p12. The results suggest that the isodisomy was not complete because of a recombination event involving the proximal short arms of two maternal chromosomes. In addition, the phenotype of proportional dwarfism in the proband suggests imprinting of one or more growth-related genes on chromosome 7. 42 refs., 5 figs., 3 tabs.

  16. Expression of a gene in a 400-base-pair fragment of colicin plasmid ColE2-P9 is sufficient to cause host cell lysis.

    PubMed Central

    Pugsley, A P; Schwartz, M

    1983-01-01

    The colicin E2 immunity (ceiB) and lysis (celB) genes of colicin plasmid ColE2-P9 were cloned as a 900-base-pair insert under the control of the lac promoter in high-copy-number plasmid pUR222. Hosts carrying this plasmid were immune to colicin E2, produced increased amounts of immunity protein (molecular weight, 9,000) and two smaller proteins (molecular weights, 5,000 and 3,000), and lysed when incubated in medium containing isopropyl-beta-D-thiogalactopyranoside (IPTG). A 400-base-pair lacp-distal fragment derived from the insert in this plasmid was recloned in the same orientation into pUR222. Although hosts carrying this plasmid also lysed when grown in the presence of IPTG, they were sensitive to colicin E2 and produced increased amounts of the 5,000- and 3,000-molecular-weight proteins (but not the full-length immunity protein) when treated with IPTG. The results were consistent with the idea that expression of celB (production of the 5,000- and 3,000-molecular-weight proteins) is sufficient to cause host cell lysis in the absence of colicin production and derepression of the host cell SOS system. Images PMID:6352670

  17. Gene-Corrected Fibroblast Therapy for Recessive Dystrophic Epidermolysis Bullosa using a Self-Inactivating COL7A1 Retroviral Vector.

    PubMed

    Jacków, Joanna; Titeux, Matthias; Portier, Soizic; Charbonnier, Soëli; Ganier, Clarisse; Gaucher, Sonia; Hovnanian, Alain

    2016-07-01

    Patients with recessive dystrophic epidermolysis bullosa (RDEB) lack type VII collagen and therefore have severely impaired dermal-epidermal stability causing recurrent skin and mucosal blistering. There is currently no specific approved treatment for RDEB. We present preclinical data showing that intradermal injections of genetically corrected patient-derived RDEB fibroblasts using a Good Manufacturing Practices grade self-inactivating COL7A1 retroviral vector reverse the disease phenotype in a xenograft model in nude mice. We obtained 50% transduction efficiency in primary human RDEB fibroblasts with an average low copy number (range = 1-2) of integrated provirus. Transduced fibroblasts showed strong type VII collagen re-expression, improved adhesion properties, normal proliferative capabilities, and viability in vitro. We show that a single intradermal injection of 3 × 10(6) genetically corrected RDEB fibroblasts beneath RDEB skin equivalents grafted onto mice allows type VII collagen deposition, anchoring fibril formation at the dermal-epidermal junction, and improved dermal-epidermal adherence 2 months after treatment, supporting functional correction in vivo. Gene-corrected fibroblasts previously showed no tumorigenicity. These data show the efficacy and safety of gene-corrected fibroblast therapy using a self-inactivating vector that has now been good manufacturing grade-certified and pave the way for clinical translation to treat nonhealing wounds in RDEB patients. PMID:26994967

  18. Expression of a partially deleted gene of human type II procollagen (COL2A1) in transgenic mice produces a chondrodysplasia

    SciTech Connect

    Vandenberg, P.; Khillan, J.S.; Prockop, D.J.; Helminen, H.; Kontusaari, S.; Ala-Kokko, L. )

    1991-09-01

    A minigene version of the human gene for type II procollagen (COL2AI) was prepared that lacked a large central region containing 12 of the 52 exons and therefore 291 of the 1523 codons of the gene. The construct was modeled after sporadic in-frame deletions of collagen genes that cause synthesis of shortened pro{alpha} chains that associate with normal pro{alpha} chains and thereby cause degradation of the shortened and normal pro{alpha} chains through a process called procollagen suicide. The gene construct was used to prepare five lines of transgenic mice expressing the minigene. A large proportion of the mice expressing the minigene developed a phenotype of a chondrodysplasia with dwarfism, short and thick limbs, a short snout, a cranial bulge, a cleft palate, and delayed mineralization of bone. A number of mice died shortly after birth. Microscopic examination of cartilage revealed decreased density and organization of collagen fibrils. In cultured chondrocytes from the transgenic mice, the minigene was expressed as shortened pro{alpha}1(II) chains that were disulfide-linked to normal mouse pro{alpha}1(II) chains. Therefore, the phenotype is probably explained by depletion of the endogenous mouse type II procollagen through the phenomenon of procollagen suicide.

  19. Heterozygous mutation of c.3521C>T in COL1A1 may cause mild osteogenesis imperfecta/Ehlers-Danlos syndrome in a Chinese family

    PubMed Central

    Shi, Xianlong; Lu, Yanqin; Wang, Yanzhou; Zhang, Yu-ang; Teng, Yuanwei; Han, Wanshui; Han, Zhenzhong; Li, Tianyou; Chen, Mei; Liu, Junlong; Fang, Fengling; Dou, Conghui; Ren, Xiuzhi; Han, Jinxiang

    2015-01-01

    Summary Osteogenesis imperfecta (OI) is an inheritable connective tissue disorder with a broad clinical heterozygosis, which can be complicated by other connective tissue disorders like Ehlers-Danlos syndrome (EDS). OI/EDS are rarely documented. Most OI/EDS mutations are located in the N-anchor region of type I procollagen and predominated by glycine substitution. We identified a c.3521C>T (p.A1174V) heterozygous mutation in COL1A1 gene in a four-generation pedigree with proposed mild OI/EDS phenotype. The affected individuals had blue sclera and dentinogenesis imperfecta (DI) was uniformly absent. The OI phenotype varied from mild to moderate, with the absence of scoliosis and increased skin extensibility. Easy bruising, joint dislocations and high Beighton score were present in some affected individuals. EDS phenotype is either mild or unremarkable in some individuals. The mutation is poorly conserved and in silico prediction support the relatively mild phenotype. The molecular mechanisms of the mutation that leads to the possible OI/EDS phenotype should be further identified by biochemical analysis of N-propeptide processing and steady state collagen analysis. PMID:25674388

  20. Ehlers-Danlos Syndrome type IV: a multi-exon deletion in one of the two COL3A1 alleles affecting structure, stability, and processing of type III procollagen

    SciTech Connect

    Superti-Furga, A.; Gugler, E.; Gitzelmann, R.; Steinmann, B.

    1988-05-05

    The authors have studied a patient with severe, dominantly inherited Ehlers-Danlos syndrome type IV. The results indicate that this patient carries a deletion of 3.3 kilobase pairs in the triple helical coding domain of one of the two alleles for the pro-..cap alpha..-chains of type III collagen (COL3A1). His cultured skin fibroblasts contain equal amounts of normal length mRNA and of mRNA shortened by approximately 600 bases, and synthesize both normal and shortened pro-..cap alpha..1(III)-chains. In procollagen molecules containing one or more shortened chains, a triple helix is formed with a length of only about 780 amino acids. The mutant procollagen molecules have decreased thermal stability, are less efficiently secreted, and are not processed as their normal counterpart. The deletion in this family is the first mutation to be described in COL3A1.

  1. A Novel p. Gly630Ser Mutation of COL2A1 in a Chinese Family with Presentations of Legg–Calvé–Perthes Disease or Avascular Necrosis of the Femoral Head

    PubMed Central

    Cao, Xiang; Wu, Qiu-Yue; Li, Wei-Wei; Li, Tian-Fu; Zhang, Cui; Cui, Ying-Xia; Li, Xiao-Jun; Yin, Zhi-Min; Xia, Xin-Yi

    2014-01-01

    Objective Mutations in the type II collagen gene are associated with certain human disorders, collectively termed type II collagenopathies. They include Legg–Calvé–Perthes disease (LCPD) and avascular necrosis of the femoral head (ANFH). These two diseases are skeletal dysplasias, inherited in an autosomal dominant fashion, characterized by groin pain, dislocation of the hip and diminished joint mobility. Coxa vara and elevation of the greater trochanter of the femur comprise the typical phenotype of LCPD, but do not occur in ANFH. Lack of synthesis of type II collagen and structural defects are responsible for the major clinical outcomes, because collagen is the essential matrix protein of all connective tissues. Type II collagen, encoded by the COL2A1 gene, contains N- and C- terminal regions that are cleaved after secretion into the extracellular matrix, and the core area is composed of a triple helical (Gly–X–Y) domain. If the Gly in this specific region is replaced by other amino acids, the structure of type II collagen will be destroyed. Method Forty-five members of a four-generation family were recruited and investigated. Diagnosis was made by independent orthopedic surgeons and radiologists. A mutation of the COL2A1 gene was detected. Result In our research, we identify a heterozygous mutation (c.1888 G>A, p. Gly630Ser) in exon 29 of COL2A1 in the Gly–X–Y domain, in a Chinese family affected by LCPD and ANFH. Our findings provide significant clues to the phenotype–genotype relationships in these syndromes and may be helpful in clinical diagnosis. Furthermore, these results should assist further studies of the mechanisms underlying collagen diseases. Conclusion Our data add new variants to the repertoire of COL2A1 mutation resulting in related collagenopathies. PMID:24949742

  2. G to A substitution in 5{prime} donor splice site of introns 18 and 48 of COL1A1 gene of type I collagen results in different splicing alternatives in osteogenesis imperfecta type I cell strains

    SciTech Connect

    Willing, M.; Deschenes, S.

    1994-09-01

    We have identified a G to A substitution in the 5{prime} donor splice site of intron 18 of one COL1A1 allele in two unrelated families with osteogenesis imperfecta (OI) type I. A third OI type I family has a G to A substitution at the identical position in intron 48 of one COL1A1 allele. Both mutations abolish normal splicing and lead to reduced steady-state levels of mRNA from the mutant COL1A1 allele. The intron 18 mutation leads to both exon 18 skipping in the mRNA and to utilization of a single alternative splice site near the 3{prime} end of exon 18. The latter results in deletion of the last 8 nucleotides of exon 18 from the mRNA, a shift in the translational reading-frame, and the creation of a premature termination codon in exon 19. Of the potential alternative 5{prime} splice sites in exon 18 and intron 18, the one utilized has a surrounding nucleotide sequence which most closely resembles that of the natural splice site. Although a G to A mutation was detected at the identical position in intron 48 of one COL1A1 allele in another OI type I family, nine complex alternative splicing patterns were identified by sequence analysis of cDNA clones derived from fibroblast mRNA from this cell strain. All result in partial or complete skipping of exon 48, with in-frame deletions of portions of exons 47 and/or 49. The different patterns of RNA splicing were not explained by their sequence homology with naturally occuring 5{prime} splice sites, but rather by recombination between highly homologous exon sequences, suggesting that we may not have identified the major splicing alternative(s) in this cell strain. Both G to A mutations result in decreased production of type I collagen, the common biochemical correlate of OI type I.

  3. A{sup -2} {yields} G transition at the 3{prime} acceptor splice site of IVS17 characterizes the COL2A1 gene mutation in the original Stickler syndrome kindred

    SciTech Connect

    Williams, C.J.; Ganguly, A.; Considine, E.

    1996-06-14

    Hereditary progressive arthro-ophthalmopathy, or {open_quotes}Stickler syndrome,{close_quotes} is an autosomal dominant osteochondrodysplasia characterized by a variety of ocular and skeletal anomalies which frequently lead to retinal detachment and precocious osteoarthritis. A variety of mutations in the COL2A1 gene have been identified in {open_quotes}Stickler{close_quotes} families; in most cases studied thus far, the consequence of mutation is the premature generation of a stop codon. We report here the characterization of a COL2A1 gene mutation in the original kindred described by Stickler et al. Conformational sensitive gel electrophoresis (CSGE) was used to screen for mutations in the entire COL2A1 gene in an affected member from the kindred. A prominent heteroduplex species was noted in the polymerase chain reaction (PCR) product from a region of the gene including exons 17 to 20. Direct sequencing of PCR-amplified genomic DNA resulted in the identification of a base substitution at the A{sup -2} position of the 3{prime} splice acceptor site of IVS17. Sequencing of DNA from affected and unaffected family members confirmed that the mutation segregated with the disease phenotype. Reverse transcriptase-PCR analysis of poly A+ RNA demonstrated that the mutant allele utilized a cryptic splice site in exon 18 of the gene, eliminating 16 bp at the start of exon 18. This frameshift eventually results in a premature termination codon. These findings are the first report of a splice site mutation in classical Stickler syndrome and they provide a satisfying historical context in which to view COL2A1 mutations in this dysplasia. 25 refs., 3 figs., 1 tab.

  4. An overlapping phenotype of Osteogenesis imperfecta and Ehlers-Danlos syndrome due to a heterozygous mutation in COL1A1 and biallelic missense variants in TNXB identified by whole exome sequencing.

    PubMed

    Mackenroth, Luisa; Fischer-Zirnsak, Björn; Egerer, Johannes; Hecht, Jochen; Kallinich, Tilmann; Stenzel, Werner; Spors, Birgit; von Moers, Arpad; Mundlos, Stefan; Kornak, Uwe; Gerhold, Kerstin; Horn, Denise

    2016-04-01

    Osteogenesis imperfecta (OI) and Ehlers-Danlos syndrome (EDS) are variable genetic disorders that overlap in different ways [Cole 1993; Grahame 1999]. Here, we describe a boy presenting with severe muscular hypotonia, multiple fractures, and joint hyperflexibility, features that are compatible with mild OI and hypermobility type EDS, respectively. By whole exome sequencing, we identified both a COL1A1 mutation (c.4006-1G > A) inherited from the patient's mildly affected mother and biallelic missense variants in TNXB (p.Val1213Ile, p.Gly2592Ser). Analysis of cDNA showed that the COL1A1 splice site mutation led to intron retention causing a frameshift (p.Phe1336Valfs*72). Type 1 collagen secretion by the patient's skin fibroblasts was reduced. Immunostaining of a muscle biopsy obtained from the patient revealed a clear reduction of tenascin-X in the extracellular matrix compared to a healthy control. These findings imply that the combination of the COL1A1 mutation with the TNXB variants might cause the patient's unique phenotype. PMID:26799614

  5. Inhibitory effect of UvrD and DinG on the replication of ColE1-derived plasmids in Escherichia coli.

    PubMed

    Kang, Nalae; Choi, Eunsil; Kim, Sung-Gun; Hwang, Jihwan

    2015-09-01

    CspA has been identified as a major cold-shock protein in Escherichia coli. CspA binds to RNAs which are abnormally folded at low temperature and then acts as an RNA chaperone unfolding those RNAs. The dramatic expression of cspA at low temperature is contributed by posttranscriptional stability and robust translatability. Interestingly, when cspA mRNA encoding a premature nonsense codon was overexpressed at low temperature, cell growth was completely inhibited. This phenotype was termed LACE (the low temperature-dependent antibiotic effect of truncated cspA expression), and this lethality resulted from exclusive stalling of most ribosomes on mutant cspA mRNAs. In a previous study, we demonstrated that overexpression of the ATP-dependent DNA helicases, UvrD and DinG, suppressed the lethality and ribosome stalling caused by mutant cspA mRNA. In the present study, we attempted to elucidate how these two DNA helicases help recover normal growth under LACE condition. Interestingly, we found that UvrD and DinG appeared to have an ability to down-regulate the replication of pUC-based high copy plasmid. In plasmid copy number tests, the amount of pUC-based plasmid encoding mutant cspA was reduced by 3-10-fold when either UvrD or DinG was expressed. Through a β-galactosidase activity assay, we also confirmed that expression of the lacZα gene inserted into the pUC-based plasmid was significantly reduced due to down-regulation of plasmid replication. Our findings imply that UvrD and DinG, known as non-replicative helicases, play a novel role in the regulation of ColE1-like plasmid replication. PMID:26143370

  6. Multi-temporal monitoring of crack formation on a mountain col with low-cost unmanned aerial systems - a case study in Austria

    NASA Astrophysics Data System (ADS)

    Stary, Ulrike; Adams, Marc

    2016-04-01

    In the Tuxer Alps of Western Austria, crack formation was observed on a col at approximately 2,500 m a.s.l., in close proximity to a highly frequented hiking trail. On an area of 0.2 ha, three several meter deep cracks were identified. Here we present the results of a 3-year monitoring of this area with low-cost, unmanned aerial systems (UAS) and photogrammetric techniques. In 2013 and 2014, a custom-built fixed-wing UAS (Multiplex Mentor, wingspan 1.6 m, gross take-off weight 2.5 kg), equipped with a Sony NEX5 (16 mm prime lens, 14 MP sensor resolution) was used to map the study site. In 2015 we employed a helicopter (Thundertiger Raptor, 0.55 m blade length, gross take-off weight 2.8 kg), fitted with a GoPro2 (60° prime lens, 5 MP sensor resolution). In all three cases we recorded 1,200-2,000 images in 10-30 minutes. To georeference the images, 8-10 ground control points (GCP) were placed at the study site and measured with a Trimble GeoXT GPS device (expected accuracy 0.15 m, precision 0.3 m). Using AgiSoft's PhotoScan (v.1.1.6), Orthophotos (OP) and digital surface models (DSM) were calculated with 5 and 20 cm ground sampling distance, respectively. The visual interpretation of the OPs gave some indication, that the size of the cracks was increasing by 0.1-0.5 m (A-axis) or 0.2-0.8 m² per year. An interpretation of the DSMs was inconclusive with regard to the depth of the cracks due to shadows in the imagery and vertical or overhanging sidewalls of the cracks. Additionally the accuracy of the GCP-measurements was found to lie below the rate of change of the cracks, thus not permitting a direct calculation of difference DSM. From an operational point-of-view, the study site proved very challenging because of its exposed, high-alpine location, with high wind speeds, gusts and poor visibility hampering the UAS-missions. The monitoring campaign will continue in 2016, where the collection of additional ground-based reference data is planned (e.g. terrestrial

  7. Rare Autosomal Recessive Cardiac Valvular Form of Ehlers-Danlos Syndrome Results from Mutations in the COL1A2 Gene That Activate the Nonsense-Mediated RNA Decay Pathway

    PubMed Central

    Schwarze, Ulrike; Hata, Ryu-Ichiro; McKusick, Victor A.; Shinkai, Hiroshi; Hoyme, H. Eugene; Pyeritz, Reed E.; Byers, Peter H.

    2004-01-01

    Splice site mutations in the COL1A2 gene of type I collagen can give rise to forms of Ehlers-Danlos syndrome (EDS) because of partial or complete skipping of exon 6, as well as to mild, moderate, or lethal forms of osteogenesis imperfecta as a consequence of skipping of other exons. We identified three unrelated individuals with a rare recessively inherited form of EDS (characterized by joint hypermobility, skin hyperextensibility, and cardiac valvular defects); in two of them, COL1A2 messenger RNA (mRNA) instability results from compound heterozygosity for splice site mutations in the COL1A2 gene, and, in the third, it results from homozygosity for a nonsense codon. The splice site mutations led to use of cryptic splice donor sites, creation of a downstream premature termination codon, and extremely unstable mRNA. In the wild-type allele, the two introns (IVS11 and IVS24) in which these mutations occurred were usually spliced slowly in relation to their respective immediate upstream introns. In the mutant alleles, the upstream intron was removed, so that exon skipping could not occur. In the context of the mutation in IVS24, computer-generated folding of a short stretch of mRNA surrounding the mutation site demonstrated realignment of the relationships between the donor and acceptor sites that could facilitate use of a cryptic donor site. These findings suggest that the order of intron removal is an important variable in prediction of mutation outcome at splice sites and that folding of the nascent mRNA could be one element that contributes to determination of order of splicing. The complete absence of proα2(I) chains has the surprising effect of producing cardiac valvular disease without bone involvement. PMID:15077201

  8. Genetic linkage analysis of the Ak1, Col5a1, Epb7. 2, Fpgs, Grp78, Pbx3, and Notch1 genes in the region of mouse chromosome 2 homologous to human chromosome 9q

    SciTech Connect

    Pilz, A. ); Prohaska, R. ); Peters, J. ); Abbott, C. Western General Hospital, Edinburgh )

    1994-05-01

    The genes for adenylate kinase-1 (AK1), folyl polyglutamate synthetase (FPGS), the collagen pro[alpha]1(V) chain (COL5A1), erythrocyte protein band 7.2b (EPB72), and a proto-oncogene homeobox (PBX3) all map to the distal portion of human chromosome 9q (HSA9q) but have not previously been mapped by linkage analysis in the mouse. In this study, the authors have used two interspecific backcrosses to map the mouse homologues of each of these genes to mouse chromosome 2 (MMU2). The Ak1, Col5a1, Epb7.2, Fpgs, and Pbx3 genes were mapped with respect to the genes for Grp78, Rxra, Notch1 (the mouse homologue of TAN1), Spna2, Abl, and Hc (the mouse homologue of C5), all of which have previously been mapped by linkage analysis on MMU2 and have human homologues that map to HSA9q. Two of the reference loci for MMU2, D2Mit1 and Acra, were also mapped in the same cross to facilitate comparisons with existing maps. The consensus gene order deduced by combining data from both crosses is D2Mit1-(Dbh,Notch1)-(Col5a1,Rxra)-Spna2-Abl-(Ak1,Fpgs)-(Grp78,Pbx3)-(Epb7.2,Hc,Gsn)-Aera. These loci therefore form part of the conserved synteny between HSA9q and MMU2. 35 refs., 2 figs., 1 tab.

  9. Chemical and physical characterisation of water in an alpine permafrost area (Col d'Olen LTER site, Italian NW-Alps)

    NASA Astrophysics Data System (ADS)

    Giardino, Marco; Colombo, Nicola; Fratianni, Simona; Guenzi, Diego; Acquaotta, Fiorella; Perotti, Luigi; Freppaz, Michele; Godone, Danilo; Said Pullicino, Daniel; Martin, Maria; Viglietti, Davide; Gorra, Roberta; Mania, Ilaria; Viviano, Gaetano; Salerno, Franco; Balestrini, Raffaella

    2015-04-01

    High altitude areas in the Alps are characterised by the permafrost environment, which reacts sensitively to climate change. During the last decades several studies on alpine permafrost-related hazards have been performed, but few studies have focused on the geochemical content of the water that drains permafrost areas or outflow from rock glaciers (Williams et al., 2006; Thies et al., 2007; Krainer et al., 2011). Rock glaciers have physical and chemical influences on interflowing waters and their discharge can be highly enriched in solutes. For example, unexpected high nickel and manganese concentrations exceeding the EU limits for drinking water have been recently reported in some studies investigating rock glacier discharges (Ilyashuk et al., 2014). The present study aims to evaluate rock glacier solute fluxes into a high altitude lake in the Italian NW-Alps (Col d'Olen LTER site, Aosta Valley) in order to understand the impact of climate parameters on alpine permafrost, in particular the effects of permafrost ice melt on the water quality of mountain headwaters. This objective has been achieved through an integrated-multidisciplinary research programme involving climate analysis, rock glacier ground surface temperature investigation, water physiochemical and microbiological analyses. Nine automatic and three manned weather stations located in the surrounding areas of the rock glacier (radius: 12 km) have been used to study the relationships between climatic parameters and permafrost dynamics. Moreover, meteorological data have been collected by installing portable instruments in situ, integrated in a Mini Automatic Weather Station. To investigate the correlations between physiochemical features of water and the thermal state of the rock glacier surface, the ground temperature monitoring has been conducted. Temperature dataloggers have been buried 5/10 cm into the ground, regularly distributed on the rockglacier surface and in few surrounding sites. Total

  10. Monitoring of glacial and periglacial landforms using terrestrial laser scanning.The case of the Col des Gentianes moraine (Valais, Switzerland)

    NASA Astrophysics Data System (ADS)

    Mazotti, B.; Oppikofer, T.; Riff, F.; Lambiel, C.; Loye, A.; Jaboyedoff, M.

    2009-04-01

    Between 1977 and 1979, important civil engineering works were made on the moraine of "Col des Gentianes", which is situated 2894 meters above the sea level in the region of Mt-Fort, Valais, Switzerland. Two cableway station arrivals, a departure station to the Mt-Fort and a restaurant were built on. This moraine was formed during the last advance of the Tortin glacier during the Little Ice Age. Since 1980, the glacier has melted dramatically and the moraine is creeping. The moraine in front of the cableway departure station to the Mt-Fort sagged by 2 to 4 meters in 30 years. A large volume of ice is still present within the moraine and melting of the ice would make its stability even more precarious. Since 2007 the moraine is monitored by terrestrial laser scanning (TLS). Two TLS campaigns were made in July and October 2008 and compared to datasets acquired in 2007. The comparison of sequential TLS point clouds enabled the detection and quantification of movements in the moraine: (1) by computing oblique (shortest) or vertical differences, (2) by creating displacement vectors and (3) by profiles across the TLS point clouds. Between July and October 2008 the Tortin glacier melted by 1 to 2.5 m and the moraine creeped in direction of the glacier by 0.25 to 0.75 m. During the same period, a landslide zone has been clearly identified downslope of the cableway departure station to the Mt-Fort. Important movements between 1.5 to 5 meters were measured on this landslide through the creation of displacement vectors. This landslide scarp is delimited by 0.5 and 1 meter downward displacements in two month. Already in 2007, a less important landslide was identified and some ice had been observed in the scarp zone. The TLS permitted to analyze the distribution of these movements on the entire moraine and not only on few measurement points like given by D-GPS. The computed TLS displacement vectors are in good agreement with annual D-GPS measurements performed on this moraine

  11. Characterization of pKP-M1144, a Novel ColE1-Like Plasmid Encoding IMP-8, GES-5, and BEL-1 β-Lactamases, from a Klebsiella pneumoniae Sequence Type 252 Isolate

    PubMed Central

    Dolejska, Monika; Izdebski, Radoslaw; Dobiasova, Hana; Studentova, Vendula; Esteves, Francisco J.; Derde, Lennie P. G.; Bonten, Marc J. M.; Hrabák, Jaroslav; Gniadkowski, Marek

    2015-01-01

    IMP-8 metallo-β-lactamase was identified in Klebsiella pneumoniae sequence type 252 (ST252), isolated in a Portuguese hospital in 2009. blaIMP-8 was the first gene cassette of a novel class 3 integron, In1144, also carrying the blaGES-5, blaBEL-1, and aacA4 cassettes. In1144 was located on a ColE1-like plasmid, pKP-M1144 (12,029 bp), with a replication region of limited nucleotide similarity to those of other RNA-priming plasmids, such as pJHCMW1. In1144 and pKP-M1144 represent an interesting case of evolution of resistance determinants in Gram-negative bacteria. PMID:26033721

  12. Characterization of pKP-M1144, a Novel ColE1-Like Plasmid Encoding IMP-8, GES-5, and BEL-1 β-Lactamases, from a Klebsiella pneumoniae Sequence Type 252 Isolate.

    PubMed

    Papagiannitsis, Costas C; Dolejska, Monika; Izdebski, Radoslaw; Dobiasova, Hana; Studentova, Vendula; Esteves, Francisco J; Derde, Lennie P G; Bonten, Marc J M; Hrabák, Jaroslav; Gniadkowski, Marek

    2015-08-01

    IMP-8 metallo-β-lactamase was identified in Klebsiella pneumoniae sequence type 252 (ST252), isolated in a Portuguese hospital in 2009. blaIMP-8 was the first gene cassette of a novel class 3 integron, In1144, also carrying the blaGES-5, blaBEL-1, and aacA4 cassettes. In1144 was located on a ColE1-like plasmid, pKP-M1144 (12,029 bp), with a replication region of limited nucleotide similarity to those of other RNA-priming plasmids, such as pJHCMW1. In1144 and pKP-M1144 represent an interesting case of evolution of resistance determinants in Gram-negative bacteria. PMID:26033721

  13. A type I collagen reporter gene construct for protein engineering studies. Functional equivalence of transfected reporter COL1A1 and endogenous gene products during biosynthesis and in vitro extracellular matrix accumulation.

    PubMed

    Lamandé, S R; Bateman, J F

    1993-07-15

    A type I collagen reporter gene construct, designed to facilitate detailed analysis of the consequences of introduced structural and regulatory mutations on collagen biosynthesis and participation in the extracellular matrix, was produced by site-directed mutagenesis of the mouse COL1A1 gene. The reporter construct, pWTCI-Ile822, carried a single base change which converted the codon for amino acid 822 of the triple helix from methionine to isoleucine. This change allowed the reporter protein, [Ile822]alpha 1(I), to be distinguished from the wild-type alpha 1(I), and quantified, by its altered CNBr cleavage pattern. In mouse Mov13 cells, which synthesize no endogenous pro alpha 1(I), reporter chains associated with endogenous pro alpha 2(I), formed pepsin-stable triple helices and were secreted efficiently from the cell. The thermal stability of wild-type molecules and molecules containing the reporter [Ile822]alpha 1(I) chains was identical. The biosynthetic characteristics of wild-type and reporter chains were directly compared in stably transfected 3T6 cells. These cells did not make a distinction between reporter and endogenous alpha 1(I) chains, which were secreted from the cells at the same rate and were processed and deposited into the 3T6 cell in vitro accumulated extracellular matrix with equal efficiency. These data demonstrate that the helical sequence alteration in the reporter protein is functionally neutral and that the reporter construct, pWTCI-Ile822, is a suitable vector for the analysis of the biochemical effects of site-directed mutations in the putative COL1A1 functional domains. PMID:8343119

  14. Analysis of SLC4A11, ZEB1, LOXHD1, COL8A2 and TCF4 gene sequences in a multi-generational family with late-onset Fuchs corneal dystrophy

    PubMed Central

    TANG, HUI; ZHANG, WEN; YAN, XIN-MIN; WANG, LIN-PING; DONG, HONG; SHOU, TAO; LEI, HUO; GUO, QIANG

    2016-01-01

    The aim of the present study was to determine the genetic basis of a multi-generational family with late-onset (LO) Fuchs corneal dystrophy (FCD). Five FCD causal genes [solute carrier family 4, sodium borate transporter, member 11 (SLC4A11), zinc finger E-box binding homeobox 1 (ZEB1), lipoxygenase homology domains 1 (LOXHD1), collagen, type VIII, alpha 2 (COL8A2) and transcription factor 4 (TCF4)], previously reported to be implicated in the pathogenesis of FCD, were screened. A total of 27 variants [including 22 known single nucleotide polysingle nucleotide polymorphisms (SNPs) from the Single Nucleotide Polymorphism Database (dbSNP) and 5 variants absent from dbSNP] were detected in this FCD pedigree across the SLC4A11, ZEB1, LOXHD1 and COL8A2 genes as follows: i) 22 known SNPs from dbSNP, including 3 coding (p.R161R, p.S213S and p.T833T) and 11 non-coding variants of SLC4A11, 2 intronic SNPs of ZEB1 from dbSNP (rs220057 and rs220060), 1 intronic SNP of LOXHD1 from dbSNP (rs16939650), and 5 SNPs of COL8A2 from dbSNP (p.A35A, p.R155Q, p.L335L, p.G495G and p.T502M); and ii) 5 variants that have not been previously reported in FCD patients and that are absent from dbSNP were identified across the ZEB1 and LOXHD1 genes; these included 3 continuous indels located at the junction of the 5′-UTR and the adjacent exon 1 of ZEB1 [Indel 1 (c.-86_-53de-lins gggaggggtggaggcggaggggtGGGGGGGAAGG); Indel 2 (c.-52_-46delinsGGGAGGG); and Indel 3 (c.-45_-42delinsAGGG)], and 2 intronic variants of LOXHD1 (c.5332-126C>T and c.1809+155G>A). Apart from one intronic SNP of SLC4A11 from dbSNP (rs372201212), the pathologic consequence of which is uncertain, and 2 intron variants of LOXHD1 (c.5332-126C>T and c.1809+155G>A); the variants likely represent examples of de novo mutations. Neither of the other 24 variants provided strong evidence of pathogenesis in this FCD pedigree. An analysis of 7 SNPs in TCF4 from dbSNP, which have been associated with LO FCD in different populations

  15. Analysis of SLC4A11, ZEB1, LOXHD1, COL8A2 and TCF4 gene sequences in a multi-generational family with late-onset Fuchs corneal dystrophy.

    PubMed

    Tang, Hui; Zhang, Wen; Yan, Xin-Min; Wang, Lin-Ping; Dong, Hong; Shou, Tao; Lei, Huo; Guo, Qiang

    2016-06-01

    The aim of the present study was to determine the genetic basis of a multi-generational family with late-onset (LO) Fuchs corneal dystrophy (FCD). Five FCD causal genes [solute carrier family 4, sodium borate transporter, member 11 (SLC4A11), zinc finger E-box binding homeobox 1 (ZEB1), lipoxygenase homology domains 1 (LOXHD1), collagen, type VIII, alpha 2 (COL8A2) and transcription factor 4 (TCF4)], previously reported to be implicated in the pathogenesis of FCD, were screened. A total of 27 variants [including 22 known single nucleotide polymorphisms (SNPs) from the Single Nucleotide Polymorphism Database (dbSNP) and 5 variants absent from dbSNP] were detected in this FCD pedigree across the SLC4A11, ZEB1, LOXHD1 and COL8A2 genes as follows: i) 22 known SNPs from dbSNP, including 3 coding (p.R161R, p.S213S and p.T833T) and 11 non-coding variants of SLC4A11, 2 intronic SNPs of ZEB1 from dbSNP (rs220057 and rs220060), 1 intronic SNP of LOXHD1 from dbSNP (rs16939650), and 5 SNPs of COL8A2 from dbSNP (p.A35A, p.R155Q, p.L335L, p.G495G and p.T502M); and ii) 5 variants that have not been previously reported in FCD patients and that are absent from dbSNP were identified across the ZEB1 and LOXHD1 genes; these included 3 continuous indels located at the junction of the 5'-UTR and the adjacent exon 1 of ZEB1 [Indel 1 (c.-86_-53delins gggaggggtggaggcggaggggtGGGGGGGAAGG); Indel 2 (c.-52_-46delinsGGGAGGG); and Indel 3 (c.-45_-42delinsAGGG)], and 2 intronic variants of LOXHD1 (c.5332-126C>T and c.1809+155G>A). Apart from one intronic SNP of SLC4A11 from dbSNP (rs372201212), the pathologic consequence of which is uncertain, and 2 intron variants of LOXHD1 (c.5332-126C>T and c.1809+155G>A); the variants likely represent examples of de novo mutations. Neither of the other 24 variants provided strong evidence of pathogenesis in this FCD pedigree. An analysis of 7 SNPs in TCF4 from dbSNP, which have been associated with LO FCD in

  16. Abnormal type III collagen produced by an exon-17-skipping mutation of the COL3A1 gene in Ehlers-Danlos syndrome type IV is not incorporated into the extracellular matrix.

    PubMed Central

    Chiodo, A A; Sillence, D O; Cole, W G; Bateman, J F

    1995-01-01

    A novel heterozygous mutation of the COL3A1 gene that encodes the alpha 1(III) chains of type III collagen was identified in a family with the acrogeric form of Ehlers-Danlos syndrome type IV (EDS-IV). Cultured dermal fibroblasts produced normal and shortened alpha 1(III) chains. The triple helix of the latter chain was shortened owing to a 33 amino acid deletion of Gly-184 to Pro-216. The corresponding region of cDNA lacked 99 base pairs from nucleotides 1051 to 1149. The deletions corresponded exactly to the normal sequence encoded by exon 17 of the COL3A1 gene. The proband was heterozygous for a T to G transversion at position +2 of intron 17, which resulted in skipping of exon 17. The splicing defect was not corrected by growing the fibroblasts at 33 degrees C and no other splicing variants were identified at 33 or 37 degrees C. The affected brother had the same mutation but his unaffected mother did not. Heterotrimeric type III collagen molecules containing normal and mutant chains were retained within the cell. The mutant homotrimeric molecules were modified and secreted normally and were thermally stable. These normal characteristics of the mutant homotrimers suggested that the loss of ten Gly-Xaa-Yaa triplets (where Gly-Xaa-Yaa is a repetitive amino acid triplet structure in which Xaa and Yaa are other amino acids, proline and hydroxyproline being more common in the Yaa position) did not adversely affect the formation and stability of the triple helix or the structural requirements for secretion. However, the mutant homotrimers were not incorporated into the extracellular matrix of an in vitro model of EDS-IV dermis. The EDS-IV phenotype in this family was probably due to a deficiency in the amount of normal type III collagen available for formation of the heterotypic collagen fibrils of the extracellular matrix. Intracellular and extracellular quality-control mechanisms prevented the incorporation of heterotrimeric and homotrimeric mutant type III collagen

  17. Symmetrical P4 cleavage at cobalt half sandwich complexes [(η5-C5H5)Co(L)] (L = CO, NHC)--a computational case study on the mechanism of symmetrical P4 degradation to P2 ligands.

    PubMed

    Zarzycki, Bartosz; Bickelhaupt, F Matthias; Radius, Udo

    2013-05-28

    A full theoretical mechanistic investigation on the symmetrical cleavage of P4 at the active complex fragments [(η(5)-C5H5)Co(L)] (L = CO, (i)Pr2Im; (i)Pr2Im = 1,3-di-iso-propylimidazolin-2-ylidene), which results in the formation of the complex [{(η(5)-C5H5)Co}2(μ,η(2:2)-P2)2] 9, is presented. The overall reaction mechanism is a complex, multistep process. Rate-determining steps of the reaction sequence are two consecutive dissociations of the co-ligands L, which induce the decisive structural rearrangements of the P4 unit. The choice of the co-ligand L ( = CO, (i)Pr2Im) influences the kinetic barrier as well as the energy balance of the overall reaction path significantly. The calculations further reveal a strong influence of the entropic effect on the overall reaction. As a consequence, the energy balance of the overall formation of 9 starting from [(η(5)-C5H5)Co(CO)] precursors is almost thermoneutral and has to overcome high kinetic barriers, whereas the reaction starting from [(η(5)-C5H5)Co((i)Pr2Im)] precursors is exothermic, featuring lower transition barriers with stabilized intermediates. From the direct comparison of both reaction coordinates it seems that the entropic effect of the co-ligands is even stronger than their electronic influence, as for both investigated systems the reactions' energy profiles are almost identical up to intermediate [{(η(5)-C5H5)Co(L)}2(μ,η(2:2)-P4)] 5 (L = CO, (i)Pr2Im). After the formation of 5, the first CO dissociation step renders the reaction endothermic for L = CO, whereas in the case of (i)Pr2Im dissociation the reaction progresses exothermically. Energy decomposition analysis and fragment analysis provide a picture of the bonding mechanisms between the metal complex fragments and P4 in the case of the most significant intermediates and the final product. PMID:23515870

  18. Splicing defects in the COL3A1 gene: marked preference for 5' (donor) spice-site mutations in patients with exon-skipping mutations and Ehlers-Danlos syndrome type IV.

    PubMed Central

    Schwarze, U; Goldstein, J A; Byers, P H

    1997-01-01

    Ehlers-Danlos syndrome (EDS) type IV results from mutations in the COL3A1 gene, which encodes the constituent chains of type III procollagen. We have identified, in 33 unrelated individuals or families with EDS type IV, mutations that affect splicing, of which 30 are point mutations at splice junctions and 3 are small deletions that remove splice-junction sequences and partial exon sequences. Except for one point mutation at a donor site, which leads to partial intron inclusion, and a single base-pair substitution at an acceptor site, which gives rise to inclusion of the complete upstream intron into the mature mRNA, all mutations result in deletion of a single exon as the only splice alteration. Of the exon-skipping mutations that are due to single base substitutions, which we have identified in 28 separate individuals, only two affect the splice-acceptor site. The underrepresentation of splice acceptor-site mutations suggests that the favored consequence of 3' mutations is the use of an alternative acceptor site that creates a null allele with a premature-termination codon. The phenotypes of those mutations may differ, with respect to either their severity or their symptomatic range, from the usual presentation of EDS type IV and thus have been excluded from analysis. Images Figure 1 Figure 3 Figure 6 PMID:9399899

  19. A 19-year-old man with relapsing bilateral pneumothorax, hemoptysis, and intrapulmonary cavitary lesions diagnosed with vascular Ehlers-Danlos syndrome and a novel missense mutation in COL3A1.

    PubMed

    Abrahamsen, Bjørg J; Kulseth, Mari Ann; Paus, Benedicte

    2015-05-01

    A 19-year-old sportsman experienced a right-sided pneumothorax and hemoptysis after having had an intermittent cough and blood-tinged sputum for 2 months. A chest CT scan revealed small cavitary lesions in both lungs. The relapsing pneumothorax was treated with a chest tube twice, as well as surgically after the second relapse. Two months after surgery, the patient developed a cough, fever, and high C-reactive protein levels. At that time, large consolidations had developed in the right lung, while the left lung subsequently collapsed due to pneumothorax. The patient's physical appearance and anamnestic information led us to suspect a genetic connective tissue disease. A sequencing analysis of the COL3A1 gene identified a novel, de novo missense mutation that confirmed the diagnosis of vascular Ehlers-Danlos syndrome (EDS). This atypical presentation of vascular EDS with intrathoracic complications shows that enhanced awareness is required and demonstrates the usefulness of the genetic analyses that are clinically available for several hereditary connective tissue disorders. PMID:25940258

  20. COL18A1 is highly expressed during human adipocyte differentiation and the SNP c.1136C > T in its "frizzled" motif is associated with obesity in diabetes type 2 patients.

    PubMed

    Errera, Flavia I V; Canani, Luís H; Yeh, Erika; Kague, Erika; Armelin-Corrêa, Lucia M; Suzuki, Oscar T; Tschiedel, Balduíno; Silva, Maria Elizabeth R; Sertié, Andréa L; Passos-Bueno, Maria Rita

    2008-03-01

    Collagen XVIII can generate two fragments, NC11-728 containing a frizzled motif which possibly acts in Wnt signaling and Endostatin, which is cleaved from the NC1 and is a potent inhibitor of angiogenesis. Collagen XVIII and Wnt signaling have recently been associated with adipogenic differentiation and obesity in some animal models, but not in humans. In the present report, we have shown that COL18A1 expression increases during human adipogenic differentiation. We also tested if polymorphisms in the Frizzled (c.1136C>T; Thr379Met) and Endostatin (c.4349G>A; Asp1437Asn) regions contribute towards susceptibility to obesity in patients with type 2 diabetes (113 obese, BMI > or =30; 232 non-obese, BMI < 30) of European ancestry. No evidence of association was observed between the allele c.4349G>A and obesity, but we observed a significantly higher frequency of homozygotes c.1136TT in obese (19.5%) than in non-obese individuals (10.9%) [P = 0.02; OR = 2.0 (95%CI: 1.07-3.73)], suggesting that the allele c.1136T is associated to obesity in a recessive model. This genotype, after controlling for cholesterol, LDL cholesterol, and triglycerides, was independently associated with obesity (P = 0.048), and increases the chance of obesity in 2.8 times. Therefore, our data suggest the involvement of collagen XVIII in human adipogenesis and susceptibility to obesity. PMID:18345385

  1. A point mutation in an intronic branch site results in aberrant splicing of COL5A1 and in Ehlers-Danlos syndrome type II in two British families.

    PubMed

    Burrows, N P; Nicholls, A C; Richards, A J; Luccarini, C; Harrison, J B; Yates, J R; Pope, F M

    1998-08-01

    Ehlers-Danlos syndrome (EDS) is a heterogeneous group of connective-tissue disorders characterized by skin fragility, joint laxity, and skeletal deformities. Type V collagen appears to have a causal role in EDS types I and II, which show phenotypic overlap and may sometimes be allelic. Type V collagen can exist as a heterotrimer, [alpha1(V)]2alpha2(V), and it both coassembles with and regulates type I collagen-fibril diameter. Using an intragenic COL5A1 polymorphism, we have demonstrated linkage, at zero recombination, to the same allele in two large British EDS type II families (LOD scores 4.1 and 4.3). Affected members from each family were heterozygous for a point mutation in intron 32 (IVS32:T-25G), causing the 45-bp exon 33 to be lost from the mRNA in approximately 60% of transcripts from the mutant gene. This mutation lies only 2 bp upstream of a highly conserved adenosine in the consensus branch-site sequence, which is required for lariat formation. Although both families shared the same marker allele, we have been unable to identify a common genealogy. This is the first description of a mutation at the lariat branch site, which plays a pivotal role in the splicing mechanism, in a collagen gene. Very probably, the resulting in-frame exon skip has a dominant-negative effect due to incorporation of the mutant proalpha chain into the triple-helical molecule. These findings further confirm the importance of type V collagen in the causation of EDS type II, and the novel collagen mutation indicates the importance of the lariat branch site in splicing. PMID:9683580

  2. Abnormal dentin structure in two novel gene mutations [COL1A1, Arg134Cys] and [ADAMTS2, Trp795-to-ter] causing rare type I collagen disorders.

    PubMed

    De Coster, P J; Cornelissen, M; De Paepe, A; Martens, L C; Vral, A

    2007-02-01

    Histological and ultrastructural observations of dentin of two patients affected with rare types of type I collagen disorders are presented. In the first case, a homozygous nonsense mutation in ADAMTS2 (substitution of a codon for tryptophan by a stopcodon) causes type VIIC Ehlers-Danlos syndrome (EDS) with multiple tooth agenesis and focal dysplastic dentin defects. In the second case, a missense mutation in COL1A1 (substitution of arginine by cysteine) results in a type I EDS phenotype with clinically normal-appearing dentition. Tooth samples are investigated by using light microscopy (LM), transmission electron microscopy (TEM) and immunostaining for types I and III collagen, and tenascin. These are compared with samples from patients with types III and IV osteogenesis imperfecta (OI) in association with dentinogenesis imperfecta (DI), showing a consistently abnormal appearance of the dentin in all specimens, with variations being primarily those of degree of change. Similarities in histological changes include the alternating presence of normal and severe pathological areas in primary and secondary dentin, the latter being characterized by large canal-like structures in atubular areas. Ultrastructural evidence of pathological dentinogenesis include abnormal distribution, size and organization of collagen fibers, which may also be found in clinically unaffected teeth. The histological and ultrastructural changes seen can be explained on the basis of odontoblast dysfunction which may be secondary to the collagen defect, interfering with different levels of odontoblast cell function and intercellular communication. These observations on (ultra)structural dentin defects associated with the two novel gene mutations are the first ever reported. PMID:17118335

  3. A 18-yr long (1993-2011) snow and meteorological dataset from a mid-altitude mountain site (Col de Porte, France, 1325 m alt.) for driving and evaluating snowpack models

    NASA Astrophysics Data System (ADS)

    Morin, S.; Lejeune, Y.; Lesaffre, B.; Panel, J.-M.; Poncet, D.; David, P.; Sudul, M.

    2012-02-01

    A quality-controlled snow and meteorological dataset spanning the period 1 August 1993-31 July 2011 is presented, originating from the experimental station Col de Porte (1325 m altitude, Chartreuse range, France). Emphasis is placed on meteorological data relevant to the observation and modelling of the seasonal snowpack. In-situ driving data, at the hourly resolution, consist in measurements of air temperature, relative humidity, wind speed, incoming short-wave and long-wave radiation, precipitation rate partitioned between snow- and rainfall, with a focus on the snow-dominated season. Meteorological data for the three summer months (generally from 10 June to 20 September), when the continuity of the field record is not warranted, are taken from a local meteorological reanalysis (SAFRAN), in order to provide a continuous and consistent gap-free record. Evaluation data are provided at the daily (snow depth, snow water equivalent, runoff and albedo) and hourly (snow depth, albedo, runoff, surface temperature, soil temperature) time resolution. Internal snowpack information are provided from weekly manual snowpit observations (mostly consisting in penetration resistance, snow type, snow temperature and density profiles) and from a hourly record of temperature and height of vertically free "settling" disks. This dataset has been partially used in the past to assist in developing snowpack model and is presented here comprehensively for the purpose of multi-year model performance assessment. The data is placed on the PANGAEA repository (http://doi.pangaea.de/10.1594/PANGAEA.774249) as well as on the public ftp server ftp://ftp-cnrm.meteo.fr/pub-cencdp/.

  4. An 18-yr long (1993-2011) snow and meteorological dataset from a mid-altitude mountain site (Col de Porte, France, 1325 m alt.) for driving and evaluating snowpack models

    NASA Astrophysics Data System (ADS)

    Morin, S.; Lejeune, Y.; Lesaffre, B.; Panel, J.-M.; Poncet, D.; David, P.; Sudul, M.

    2012-07-01

    A quality-controlled snow and meteorological dataset spanning the period 1 August 1993-31 July 2011 is presented, originating from the experimental station Col de Porte (1325 m altitude, Chartreuse range, France). Emphasis is placed on meteorological data relevant to the observation and modelling of the seasonal snowpack. In-situ driving data, at the hourly resolution, consist of measurements of air temperature, relative humidity, windspeed, incoming short-wave and long-wave radiation, precipitation rate partitioned between snow- and rainfall, with a focus on the snow-dominated season. Meteorological data for the three summer months (generally from 10 June to 20 September), when the continuity of the field record is not warranted, are taken from a local meteorological reanalysis (SAFRAN), in order to provide a continuous and consistent gap-free record. Data relevant to snowpack properties are provided at the daily (snow depth, snow water equivalent, runoff and albedo) and hourly (snow depth, albedo, runoff, surface temperature, soil temperature) time resolution. Internal snowpack information is provided from weekly manual snowpit observations (mostly consisting in penetration resistance, snow type, snow temperature and density profiles) and from a hourly record of temperature and height of vertically free ''settling'' disks. This dataset has been partially used in the past to assist in developing snowpack models and is presented here comprehensively for the purpose of multi-year model performance assessment. The data is placed on the PANGAEA repository (http://dx.doi.org/10.1594/PANGAEA.774249) as well as on the public ftp server ftp://ftp-cnrm.meteo.fr/pub-cencdp/.

  5. Reflections upon University Retirement: With Thanks and Apologies to James Joyce

    ERIC Educational Resources Information Center

    Euster, Gerald L.

    2004-01-01

    This article examines (1) the elimination of mandatory retirement in American higher education, (2) research related to retirement decisions of university faculty, and (3) the author's thoughts and feelings before and following retirement from academia. An interior monologue, stream-of-consciousness method is used to reflect the author's movement…

  6. Invited address: James Joyce, Alice in Wonderland, The Rolling Stones, and criminal careers.

    PubMed

    Piquero, Alex R

    2011-07-01

    The study of criminal careers generally, and patterns of continuity and change in criminal offending in particular, has been a long-standing interest to social scientists across many disciplines. This article provides readers with an overview of this line of research. After an introduction to the criminal career perspective, the article presents several 'facts' that have emerged from criminal career studies. This material segues into a discussion of theories based on criminal careers research as well as a related discussion of the emerging methods and trends in the area. The article closes with some observations about public policy with respect to criminal careers knowledge and identifies some neglected research needs. A key summary conclusion is that the processes associated with continuity and change are not mutually exclusive, but instead are important and complimentary aspects of criminal careers research. PMID:21643670

  7. Using Joyce/Showers Teaching Models in the High School Classroom.

    ERIC Educational Resources Information Center

    Moeschl, Sue H.; Costello, Ronald W.

    1988-01-01

    Discusses the use of three teaching models in an Indianapolis high school: concept attainment, Taba (inductive thinking), and synectics. Describes each model, its use in the classroom, and the effect on the classroom climate. (MS)

  8. Invited Address: James Joyce, Alice in Wonderland, the Rolling Stones, and Criminal Careers

    ERIC Educational Resources Information Center

    Piquero, Alex R.

    2011-01-01

    The study of criminal careers generally, and patterns of continuity and change in criminal offending in particular, has been a long-standing interest to social scientists across many disciplines. This article provides readers with an overview of this line of research. After an introduction to the criminal career perspective, the article presents…

  9. Are You Experienced? Teaching and Reading Joy(ce) through the Body

    ERIC Educational Resources Information Center

    Utell, Janine

    2007-01-01

    Teacher authority is wielded in such a way that it is no longer "the" authority. Authority is used to open a space for students to find their own authority through speaking and writing--through voice. This process, so central to the feminist classroom, leads to the goals of feminist pedagogy as articulated by Robbin Crabtree and David Alan Sapp:…

  10. Acknowledging a Problem Is the First Step to Solving It: A Response to Joyce Annunziata.

    ERIC Educational Resources Information Center

    Fossey, Richard

    1999-01-01

    Acknowledges the exemplary hiring practices of the Miami-Dade County (Florida) School District, but reiterates that throughout the United States, many professional educators are hired who lack the skills, compassion, and creativity to be in charge of children's education. (Author/SLD)

  11. Intérêt de l'inspection visuelle à l'acide acétique et au soluté de Lugol avec colposcope dans le dépistage des lésions du col utérin au Gabon

    PubMed Central

    Mpiga, Édith; Ivanga, Mahinè; Koumakpayi, Ismaël Hervé; Engohan-Aloghe, Corinne; Ankély, Junie Chansi; Belembaogo, Ernest; Meye, Jean-François

    2015-01-01

    Introduction Au Gabon, le dépistage des lésions précancéreuses et cancéreuses du col de l'utérus n'est pas systématique. La vulgarisation de ce dépistage suppose l'utilisation d'un test efficace et adapté aux réalités locales. Méthodes L'objectif de cette étude était de comparer les performances du frottis cervico-vaginal (FCV) conventionnel et de l'inspection visuelle à l'acide acétique (IVA) et au soluté de Lugol (IVL) couplée à la colposcopie, dans la détection des lésions du col utérin au Gabon. Des tests IVA/IVL et FCV ont été effectués chez 309 femmes gabonaises âgées de 18 à 75 ans. Des biopsies ont été réalisées en cas de résultat positif. Résultats 5 cancers épidermoïdes (1,6%) et 4 lésions précancéreuses (1,3%) ont été confirmées par l'histologie. L'IVA/IVL a présenté une meilleure sensibilité (100%) que le FCV (89%). Toutefois, le FCV est apparu plus spécifique (100% versus 92%). Avec une valeur prédictive (VP) négative de 100%, l'IVA/IVL a permis d'exclure avec certitude la présence de cancer du col lorsque le résultat était négatif, contrairement au FCV (92%). L'IVA/IVL et le FCV ont présenté des VP positives respectivement de 90% et 100%. Conclusion Cette étude montre que l'IVA/IVL avec colposcope couplée à l'histologie en cas de résultat positif apparaît plus performante que le FCV. PMID:26893799

  12. A 5' splice site mutation affecting the pre-mRNA splicing of two upstream exons in the collagen COL1A1 gene. Exon 8 skipping and altered definition of exon 7 generates truncated pro alpha 1(I) chains with a non-collagenous insertion destabilizing the triple helix.

    PubMed Central

    Bateman, J F; Chan, D; Moeller, I; Hannagan, M; Cole, W G

    1994-01-01

    A heterozygous de novo G to A point mutation in intron 8 at the +5 position of the splice donor site of the gene for the pro alpha 1(I) chain of type I procollagen, COL1A1, was defined in a patient with type IV osteogenesis imperfecta. The splice donor site mutation resulted not only in the skipping of the upstream exon 8 but also unexpectedly had the secondary effect of activating a cryptic splice site in the next upstream intron, intron 7, leading to re-definition of the 3' limit of exon 7. These pre-mRNA splicing aberrations cause the deletion of exon 8 sequences from the mature mRNA and the inclusion of 96 bp of intron 7 sequence. Since the mis-splicing of the mutant allele product resulted in the maintenance of the correct codon reading frame, the resultant pro alpha 1(I) chain contained a short non-collagenous 32-amino-acid sequence insertion within the repetitive Gly-Xaa-Yaa collagen sequence motif. At the protein level, the mutant alpha 1(I) chain was revealed by digestion with pepsin, which cleaved the mutant procollagen within the protease-sensitive non-collagenous insertion, producing a truncated alpha 1(I). This protease sensitivity demonstrated the structural distortion to the helical structure caused by the insertion. In long-term culture with ascorbic acid, which stimulates the formation of a mature crosslinked collagen matrix, and in tissues, there was no evidence of the mutant chain, suggesting that during matrix formation the mutant chain was unable to stably incorporated into the matrix and was degraded proteolytically. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:7945197

  13. Misura dell'azimut della Piramide Cestia col Sole

    NASA Astrophysics Data System (ADS)

    Sigismondi, Costantino

    2016-05-01

    The Pyramid of Caius Cestius in Rome has been built in 330 days in 12 b.C. The instants when the Sun rays are grazing to the East face of the Pyramid have been observed in different days, writing a table of solar azimut and altitudes. Extrapolating for zero altitude, the azimut 201.8°±1.4° of the Pyramid is obtained; it corresponds to the azimut 204.27° of the Sun when the shadow of the lightning rod divides in two equal parts the North face.

  14. Fotometria col filtro LPR alla nova V5668 SGR

    NASA Astrophysics Data System (ADS)

    Sigismondi, Costantino

    2016-09-01

    The nova SGR 2015 no. 2 or V5668 SGR was visually observed with the LPR filter to evaluate the OIII emission. The contributions of B, V, R and Visual components to the LPR magnitude are modeled and compared with the observations in scotopic vision.

  15. Evaluation des résultats après traitement des lésions intra épithéliales du col utérin par la cryothérapie: étude préliminaire au Centre Hospitalier Universitaire de Yaoundé: A propos de 21 cas

    PubMed Central

    Ndoua, Claude Cyrille Noa; Tebeu, Pierre Marie; Kemfang, Jean Dupont; Kasia, Jean Marie

    2015-01-01

    Nous rapportons les résultats d'une série de 21 cas de prise en charge par cryothérapie de lésions intra-épithéliales cervicales au Centre Hospitalier et Universitaire (CHU) de Yaoundé. Notre objectif principal était d’évaluer les résultats préliminaires de la prise en charge des lésions précancéreuses éligibles pour la cryothérapie. Il s'agissait d'une étude transversale descriptive qui s'est étalée sur 24 mois. Etaient inclus dans l’étude toutes les femmes traitées par cryothérapie. Nous avons exclu les patientes traitées par une autre méthode, les patientes perdues de vue et les dossiers incomplets. Le statut cervical a été déterminé à 6 semaines, 6 mois et 12 mois. Les complications précoces et tardives ont également été répertoriées. Au total 95.2% des lésions étaient cicatrisées à 6 semaines. A 6 mois, toutes les lésions avaient disparu et au 12ème mois, la guérison était effective chez 95.2% des patientes. Les saignements et l'hydrorrhée étaient les principales complications tardives avec des fréquences respectives de 66.7% et 95.2%. Aucun cas de sténose cervicale n'a été répertorié. La cryothérapie peut être utilisée comme méthode de traitement pour des lésions précancéreuses du col. PMID:26140068

  16. Community Responses to School Reform in Chicago: Opportunities for Local Stakeholder Engagement. A Report by Public Agenda for the Joyce Foundation

    ERIC Educational Resources Information Center

    Public Agenda, 2012

    2012-01-01

    This is a report on how community stakeholders, including parents, teachers, community leaders and advocates, think about current efforts by Chicago Public Schools (CPS) to "turn around" Chicago's lowest-performing schools, and their expectations for future school reform actions. It was prepared by Public Agenda, with support from…

  17. The human alpha 2(IV) collagen gene, COL4A2, is syntenic with the alpha 1(IV) gene, COL4A1, on chromosome 13.

    PubMed

    Solomon, E; Hall, V; Kurkinen, M

    1987-05-01

    We have previously assigned the gene for the alpha 1 chain of type IV collagen to chromosome 13. In this report we show that the gene coding for the second chain of this heterotrimer is on the same chromosome. This is the first example of the genes for both chains of one collagen molecule being syntenic. PMID:3674752

  18. 78 FR 47316 - National Advisory Council for Environmental Policy and Technology

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-05

    ... interpersonal, oral and written communication, and consensus-building skills. --Ability to volunteer... ] Recruitment 2014'' to joyce.mark@epa.gov . FOR FURTHER INFORMATION CONTACT: Mark Joyce, Acting Designated... AGENCY National Advisory Council for Environmental Policy and Technology AGENCY: Environmental...

  19. Art and the Joycean Artist.

    ERIC Educational Resources Information Center

    Schiralli, Martin

    1989-01-01

    Suggests that the aesthetic positions James Joyce expresses in his work have an independent intellectual value in understanding gifted childrens' aesthetic development. Discusses and critiques Joyce's theory on the psychology and philosophy of the artist. (KM)

  20. S-COL: A Copernican Turn for the Development of Flexibly Reusable Collaboration Scripts

    ERIC Educational Resources Information Center

    Wecker, Christof; Stegmann, Karsten; Bernstein, Florian; Huber, Michael J.; Kalus, Georg; Kollar, Ingo; Rathmayer, Sabine; Fischer, Frank

    2010-01-01

    Collaboration scripts are usually implemented as parts of a particular collaborative-learning platform. Therefore, scripts of demonstrated effectiveness are hardly used with learning platforms at other sites, and replication studies are rare. The approach of a platform-independent description language for scripts that allows for easy…

  1. Electron-Molecule Col1isions: Quantitative Approaches, and the Legacy of Aaron Temkin

    NASA Technical Reports Server (NTRS)

    Schneider, B.I.

    2007-01-01

    This article, on electron-molecule collisions, is dedicated to the legacy of my good friend and sometime collaborator, Aaron Temkin on his retirement from the NASA-Goddard Space Flight Center after many years of work at the highest intellectual level in the theoretical treatment of electron-atom and electron-molecule scattering. Aaron's contributions to the manner in which we think about electron-molecule collisions is clear to all of us who have worked in this field. I doubt that the great progress that has occurred in the computational treatment of such complex collision problems could have happened without these contributions. For a brief historical account, see the discussion of Temkin's contribution to electron-molecule scattering in the first article of this volume by Dr. A. K. Bhatia. In this article, I will concentrate on the application of the so called, non-adiabatic R-matrix theory, to vibrational excitation and dissociative attachment, although I will also present some results applying the Linear Algebraic and Kohn-Variational methods to vibrational excitation. As a starting point for almost all computationally effective approaches to electron-molecule collisions, is the fixed nuclei approximation. That is, one recognizes, just as one does with molecular bound states, that there is a separation of electronic(fast) and nuclear(s1ow) degrees of freedom. This separation makes it possible to "freeze" the nuclei in space, calculate the collision parameters for the frozen molecule and then, somehow to add back the vibrations and rotations. The manner in which this is done, depends on the details of the collision problem. It is the work of Aaron and a number of other researchers that has provided the guidance necessary to resolve these issues.

  2. ColDICE: A parallel Vlasov-Poisson solver using moving adaptive simplicial tessellation

    NASA Astrophysics Data System (ADS)

    Sousbie, Thierry; Colombi, Stéphane

    2016-09-01

    Resolving numerically Vlasov-Poisson equations for initially cold systems can be reduced to following the evolution of a three-dimensional sheet evolving in six-dimensional phase-space. We describe a public parallel numerical algorithm consisting in representing the phase-space sheet with a conforming, self-adaptive simplicial tessellation of which the vertices follow the Lagrangian equations of motion. The algorithm is implemented both in six- and four-dimensional phase-space. Refinement of the tessellation mesh is performed using the bisection method and a local representation of the phase-space sheet at second order relying on additional tracers created when needed at runtime. In order to preserve in the best way the Hamiltonian nature of the system, refinement is anisotropic and constrained by measurements of local Poincaré invariants. Resolution of Poisson equation is performed using the fast Fourier method on a regular rectangular grid, similarly to particle in cells codes. To compute the density projected onto this grid, the intersection of the tessellation and the grid is calculated using the method of Franklin and Kankanhalli [65-67] generalised to linear order. As preliminary tests of the code, we study in four dimensional phase-space the evolution of an initially small patch in a chaotic potential and the cosmological collapse of a fluctuation composed of two sinusoidal waves. We also perform a "warm" dark matter simulation in six-dimensional phase-space that we use to check the parallel scaling of the code.

  3. X-24B with Test Pilot Lt. Col. Michael V. Love

    NASA Technical Reports Server (NTRS)

    1976-01-01

    This photo shows Air Force Lieutenant Colonel Michael V. Love in front of the X-24B lifting body research vehicle at Edwards Air Force Base in 1976. Love was assigned as a project pilot on the joint NASA-USAF X-24B Lifting Body flight test program at the NASA Flight Research Center. He made a total of 12 flights in the plane from October 4, 1973 until July 15, 1975. Love flew it to a speed of Mach 1.76 on October 25, 1974, a record for the X-24B. Love attended the USAF Test Pilot School and remained as an instructor there from 1969 through 1971. He was a test pilot at Edwards when assigned to fly to the X-24B. Love was a combat veteran of Vietnam and was awarded the Distinguished Flying Cross with two Oak Leaf clusters. Love perished while attempting an emergency landing in an RF-4C on March 1, 1976 - less than a month after this photo was taken. The X-24B was the last aircraft to fly in the Dryden Flight Research Center's manned lifting body program. The X-24 was one of a group of lifting bodies flown by the NASA Flight Research Center (now Dryden Flight Research Center), Edwards, California, in a joint program with the U.S. Air Force at Edwards Air Force Base from 1963 to 1975. The lifting bodies were used to demonstrate the ability of pilots to maneuver and safely land wingless vehicles designed to fly back to Earth from space and be landed like an airplane at a predetermined site. Lifting bodies' aerodynamic lift, essential to flight in the atmosphere, was obtained from their shape. The addition of fins and control surfaces allowed the pilots to stabilize and control the vehicles and regulate their flight paths. Built by Martin Aircraft Company, Maryland, for the U.S. Air Force, the X-24A was a bulbous vehicle shaped like a teardrop with three vertical fins at the rear for directional control. It weighed 6,270 pounds, was 24.5 feet long and 11.5 feet wide (measuring just the fuselage, not the distance between the tips of the outboard fins). Its first unpowered glide flight was on April 17, 1969, with Air Force Maj. Jerauld Gentry at the controls. Gentry also piloted its first powered flight on March 19, 1970. The X-24A was flown 28 times in the program that, like the HL-10, validated the concept that a Space Shuttle vehicle could be landed unpowered. The fastest speed achieved by the X-24A was 1,036 miles per hour (mph--Mach 1.6). Its maximum altitude was 71,400 feet. It was powered by an XLR-11 rocket engine with a maximum theoretical vacuum thrust of 8,480 pounds. The X-24A was later modified into the X-24B. The bulbous shape of the X-24A was converted into a 'flying flatiron' shape with a rounded top, flat bottom, and double delta platform that ended in a pointed nose. The X-24B demonstrated that accurate unpowered reentry vehicle landings were operationally feasible. Top speed achieved by the X-24B was 1,164 mph and the highest altitude it reached was 74,130 feet. The vehicle is on display at the Air Force Museum, Wright-Patterson Air Force Base, Ohio. The pilot on the last powered flight of the X-24B was Bill Dana, who also flew the last X-15 flight about seven years earlier. The X-24A shape was later borrowed for the X-38 Crew Return Vehicle (CRV) technology demonstrator for the International Space Station. The X-24B is on public display at the Air Force Museum, Wright-Patterson AFB, Ohio.

  4. Brig. Gen. Richard F. Abel and Col. Natan J. Lindsay answering questions

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Brigadier General Richard F. Abel, right, director of public affairs for the Air Force, and Colonel Nathan J. Lindsay of the USAF's space division, answer questions concerning STS-4 during a press conference at JSC on May 20, 1982.

  5. Keynote speaker Col. Fitch talks to employee audience at Super Safety and Health Day at KSC.

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Capt. Dennis E. Fitch, a consultant and former pilot instructor with United Airlines, addresses an audience of KSC employees to kick off Super Safety and Health Day at KSC. Fitch related his tale of the catastrophic engine failure in UAL flight 232, which crash landed in Iowa in 1989, and the teamwork that contributed to his survival and the lives of 183 other passengers. For the second time Kennedy Space Center dedicated an entire day to safety and health. Most normal work activities were suspended to allow personnel to attend Super Safety and Health Day activities. The theme, 'Safety and Health Go Hand in Hand,' emphasized KSC's commitment to place the safety and health of the public, astronauts, employees and space-related resources first and foremost. Events also included a panel session about related issues, vendor exhibits, and safety training in work groups. The keynote address and panel session were also broadcast internally over NASA television.

  6. Fine structure and functional comments of mouthparts in Platypus cylindrus (Col., Curculionidae: Platypodinae).

    PubMed

    Belhoucine, Latifa; Bouhraoua, Rachid T; Prats, Eva; Pulade-Villar, Juli

    2013-02-01

    Oak pinhole borer, Platypus cylindrus is seen in recent years as one of the biggest enemies directly involved in the observed decline of cork oak in Mediterranean forests with all the economic implications. As an ambrosia beetle, it has developed its effective drilling mouthpart enough to make tunnels in hardwood of the tree. The fine structural aspects of the mouthpart using the field emission scanning electron microscopy are analyzed about 23 adults collected in galleries of infested cork oak trees (Quercus suber) in a littoral forest of northwest Algeria. These adults are preserved in alcohol 70%, cleaned and coated with gold. The mouthparts of this beetle consist commonly of a labrum, a pair of mandibles, a pair of maxillae and the labium but with adapted structure to excavate galleries in the hardwood. In this role is also involved the first pair of legs. The function that present the different structures related to the construction of the tunnels is discussed. Both of maxillary and labial palpi direct the food to the mouth and hold it while the mandibles chew the food. The distal ends of these palpi are flattened and have shovel-like setae. Females have larger maxillary palpi than males and this is related to the particular biology of each sex. PMID:23182681

  7. Scientific discovery, peak experiences and the Col-oh-nell Flastratus! phenomenon.

    PubMed

    Charlton, Bruce G

    2007-01-01

    Once I had a bizarre dream in which I was vouchsafed a secret which would ensure my wealth and success. It was the title for a comic novel; one supposedly so funny that it would guarantee classic status for any book: Oh Colonel Flastratus! The distinctive feature about my dream was its quality of profound significance, which felt akin to the Eureka moment of a scientific discovery. This led me to question whether the 'peak experience' (PE) of scientific discovery might be as delusional as my dream. On the one hand, euphoric elation attached to a discovery does not guarantee that insights objective truth - implications must be spelled-out and checked. The easy induction of pseudo-profound insights by intoxicants serves as a warning of the potential pitfalls. An arbitrary object becomes labeled with an obscure sense of delight and personal relevance in a process that could be termed the Colonel Flastratus! phenomenon. But neither are peak experiences irrelevant. A scientific PE is some kind of personal guarantee of the subjective truth of an insight - a signal that states: 'This is high quality stuff, by your standards. Do not ignore it, do not forget it, try to understand it'. Peak experiences in science could therefore be considered the result of a 'significance alarm' going off in the brain and their objective value depends on the specialized cognitive quality of that specific brain. So scientists may be correct to take peak experiences seriously. Perhaps the best approach is to regard the scientific PE as a signal from the self to the self, a subjectively evaluated and auto-administered emotional reward for good thinking. PMID:17543473

  8. Complete Genome Sequence of Porcine Epidemic Diarrhea Virus Strain COL/Cundinamarca/2014 from Colombia

    PubMed Central

    Jarvis, Matthew C.; Lam, Ham Ching; Rovira, Albert

    2016-01-01

    Porcine epidemic diarrhea virus (PEDV) has been found throughout Europe and Asia, and has emerged in North and South America. A whole genome sequence was obtained from a paraffin-embedded tissue sample from the Instituto Colombiano Agropecuario (ICA), Colombia through Next Generation Sequencing techniques to further understand the evolution of PEDV. PMID:27103712

  9. Characterization and distribution of ColE1-like kanamycin-resistance plasmids in Salmonella enterica from food animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Antimicrobial resistant foodborne pathogens cause public health concerns and multi-drug resistant (MDR) pathogens present difficulties when treatment is warranted. Large plasmids are responsible for the majority of the MDR and subsequently, the focus of most research. Previous studies sh...

  10. The Placenta in Monoclea forsteri Hook. and Treubia lacunosa (Col.) Prosk: Insights into Placental Evolution in Liverworts

    PubMed Central

    CARAFA, A.; DUCKETT, J. G.; LIGRONE, R.

    2003-01-01

    Placental morphology is remarkably diverse between major bryophyte groups, especially with regard to the presence and distribution of transfer cells in the sporophyte and gametophyte. In contrast, with the exception of metzgerialean liverworts, placental morphology is highly conserved within major bryophyte groups. Here we examine the ultrastructure of the placenta in Monoclea forsteri and Treubia lacunosa, basal members of the marchantialean and metzgerialean liverwort lineages, respectively. In both species several layers of transfer cells are found on both sides of the placenta, with sporophytic transfer cells exhibiting prominent wall labyrinths. Consistent with previous reports of a similar placenta in other putatively basal and isolated liverwort genera such as Fossombronia, Haplomitrium, Blasia and Sphaerocarpos, this finding suggests that this type of placenta represents the plesiomorphic (primitive) condition in liverworts. Distinctive ultrastructural features of placental cells in Monoclea include branched plasmodesmata in the sporophyte and prominent arrays of smooth endoplasmic reticulum, seemingly active in secretion in the gametophyte. These arrays contain a core of narrow tubules interconnected by electron‐opaque rods, structures with no precedent in plants. Analysis of the distribution of different types of placenta in major bryophyte groups provides valuable insights into their inter‐relationships and possible phylogeny. PMID:12876192

  11. 77 FR 64718 - Safety Zone; Steam Ship Col. James M. Schoonmaker Relocation Project, Maumee River, Toledo, OH

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-23

    ... Schoonmaker'). This temporary safety zone will encompass all waters of the Maumee River within a 100 yard... Security FR Federal Register NPRM Notice of Proposed Rulemaking A. Regulatory History and Information The...) Location. The following area is a temporary safety zone: all waters of the Maumee River within a 100...

  12. Preclinical Alterations in the Serum of COL(IV)A3(-)/(-) Mice as Early Biomarkers of Alport Syndrome.

    PubMed

    Muckova, Petra; Wendler, Sindy; Rubel, Diana; Büchler, Rita; Alert, Mandy; Gross, Oliver; Rhode, Heidrun

    2015-12-01

    The efficiency of the inhibition of the angiotensin converting enzyme, the most widely used therapy for the Alport syndrome, depends on the onset of the therapy-the earlier the better. Hence, early progressive biomarkers are urgently required to allow for preclinical diagnosis, an early start of possible therapy as well as the monitoring of this therapy. In the present study, an improved comprehensive and precise proteomic approach has been applied to the serum of juvenile Alport-mice, nontreated and treated, and wild-type controls of various ages to search for biomarkers. With a total of 2542 stringently altered proteins, the serum composition clearly shows a dependency on age, that is, stage, and therapy. Initially, the serum constituents indicate an enhanced extracellular matrix remodeling, cell damage, and the production of particular acute phase proteins. A panel of 15 potential biomarker candidates has been identified. In later stages, renal filtration failure and systemic acute phase reaction determine the composition of the serum; an effect that is well-known for manifested human Alport syndrome. With a small number of mouse urine samples, for example, the proteomic results for gelsolin could be verified using ELISA. Once verified in man, these early biomarkers would allow for a sensitive and specific diagnosis of the Alport syndrome in children as well as facilitate the monitoring of a possible therapy. PMID:26487288

  13. Cytotoxic activity and inhibition of tumor cell invasion by derivatives of a chemically modified tetracycline CMT-3 (COL-3).

    PubMed

    Lokeshwar, B L; Escatel, E; Zhu, B

    2001-02-01

    Tetracyclines such as chlortetracycline and doxycycline with antimicrobial activity were reported to possess cytostatic and cytotoxic activity against mammalian tumor cells, often at high doses. Non-antimicrobial chemically modified tetracyclines (CMTs), with limited systemic toxicity but with significant tumor cell toxicity and antimetastatic activity, are attractive for long term treatment for cancer. We recently reported one such CMT, 6-deoxy,6-demethyl 4-dedimethylamino tetracycline (CMT-3) is a potent anti-tumor and anti-metastatic drug. Here we report on the anti-cell proliferation and anti-invasive activity of five nitro derivatives of CMT-3 (CMT-3N). All the five CMT-3Ns (CMT-302, CMT-303, CMT-306, CMT-308 and CMT-316) inhibited in vitro cell proliferation of prostate cancer cells. The 50% growth inhibition concentration (IC(50)) of CMT-3Ns was similar to that of CMT-3. Although CMT-3 was by far the most potent anti-cell proliferation drug, all CMT-3Ns except CMT-303 and CMT-308 had similar anti-cell proliferation activity (IC(50): 2.5 -5.7 microg/ml). IC(50)s for CMT-303 and CMT-308 were approximately 8.1 and -12.4 microg/ml, respectively. Activity against tumor cell invasion was tested in vitro using the Matrigel invasion assay. All CMT-3Ns had similar anti- invasive activity. While cytotoxic activity of CMT-3 was strongly associated with cell death-effector caspase activation, mitochondrial permeablization and apoptosis, the CMT-3Ns weakly induced apoptosis and did not activate Caspase-3. However, the CMT-3Ns were able to induce mitochondrial permeabilization. This dichotomous mechanism of cytotoxic activity of CMTs may have significance in their selection for clinical application. PMID:11172682

  14. Nuclear sequestration of COL1A1 mRNA transcript associated with type I osteogenesis imperfecta (OI)

    SciTech Connect

    Primorac, D.; Stover, M.L.; McKinstry, M.B.

    1994-09-01

    Previously we identified an OI type I patient with a splice donor mutation that resulted in intron 26 retention instead of exon skipping and sequestration of normal levels of the mutant transcript in the nuclear compartment. Intron retention was consistent with the exon definition hypothesis for splice site selection since the size of the exon-intron-exon unit was less than 300 bp. Furthermore, the retained intron contained in-frame stop codons which is thought to cause the mutant RNA to remain within the nucleus rather than appearing in the cytoplasm. To test these hypotheses, genomic fragments containing the normal sequence or the donor mutation were cloned into a collagen minigene and expressed in stably tansfected NIH 3T3 cells. None of the modifications to the normal intron altered the level of RNA that accumulated in the cytoplasm, as expected. However none of the modifications to the mutant intron allowed accumulation of normal levels of mRNA in the cytoplasm. Moreover, in contrast to our findings in the patient`s cells only low levels of mutant transcript were found in the nucleus; a fraction of the transcript did appear in the cytoplasm which had spliced the mutant donor site correctly. Nuclear run-on experiments demonstrated equal levels of transcription from each transgene. Expression of another donor mutation known to cause in-frame exon skipping in OI type IV was accurately reproduced in the minigene in transfected 3T3 cells. Our experience suggests that either mechanism can lead to formation of a null allele possibly related to the type of splicing events surrounding the potential stop codons. Understanding the rules governing inactivation of a collagen RNA transcript may be important in designing a strategy to inactivate a dominate negative mutation associated with the more severe forms of OI.

  15. 75 FR 15673 - Notice of Request for Extension of Approval of an Information Collection; Export Health...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-30

    ... accordance with the Paperwork Reduction Act of 1995, this notice announces the Animal and Plant Health... United States, contact Dr. Joyce Bowling- Heyward, Assistant Director, Technical Trade Services...

  16. A Consumer's Guide to High School History Textbooks

    ERIC Educational Resources Information Center

    Ravitch, Diane

    2004-01-01

    This document reviews a dozen history textbooks. The following American History textbooks are reviewed: (1) Joyce Appleby, Alan Brinkley, and James M. McPherson, The American Journey: Building a Nation (Glencoe, 2003), and Joyce Appleby, Alan Brinkley, Albert S. Broussard, James M. McPherson, and Donald A. Ritchie, The American Republic Since 1877…

  17. Update on Research and Leadership. Vol. 21, No. 1. Fall 2009

    ERIC Educational Resources Information Center

    Bragg, Debra D., Ed.; Taylor, Jason L., Ed.

    2009-01-01

    This edition features current research, practice, and policy related to the Joyce Foundation's Shifting Gears initiative beginning with an interview with Whitney Smith, Manager of the Employment Program at the Joyce Foundation. Julie Strawn, Center for Law and Social Policy, presents a national perspective of basic skills reform efforts similar to…

  18. 76 FR 76218 - Reports, Forms and Record Keeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-06

    ... of 1995 (44 U.S.C. 3501 et seq.), this notice announces that the Information Collection Request (ICR... submitted on or before January 5, 2012. FOR FURTHER INFORMATION CONTACT: Mike Joyce, Marketing Specialist... Administration, 1200 New Jersey Ave. SE., W52- 238, Washington, DC 20590. Mike Joyce's phone number is (202)...

  19. Shady Oaks Playscape Revisited: A Natural Playscape that Evolves over Time

    ERIC Educational Resources Information Center

    Keeler, Rusty

    2008-01-01

    Shady Oak Christian School serves children ages two and a half through second grade and in the summer provides programs for preschoolers through fifth grade. The school has an amazing home-like environment with a super go-getting director, Joyce Trigger. In this article, the author presents an interview with Joyce Trigger about her playscape and…

  20. Enhancing Students' Numeracy and Analytical Skills with the Use of Technology: A Career Preparation Approach

    ERIC Educational Resources Information Center

    Hurst, Jessica L.

    2013-01-01

    In today's competitive job market, education is increasingly touted as necessary preparation for the workplace (Joyce, Hassall, Montano, & Anes, 2006). The literature suggests that one of the most important skill sets identified by both educators and practitioners is the need for numerical competency and analytical proficiency (Joyce et…

  1. 75 FR 26255 - Change in Bank Control Notices; Acquisition of Shares of Bank or Bank Holding Companies

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-11

    ...-0291: 1. Joyce J. Eickhoff Revocable Trust, and Joyce J. Eckhoff, Trustee, both of Adrian, Minnesota; to acquire voting shares of Adrian Building Corporation, and thereby indirectly acquire voting shares of Adrian State Bank, both of Adrian, Minnesota. B. Federal Reserve Bank of San Francisco...

  2. Suspension survival mediated by PP2A-STAT3-Col XVII determines tumour initiation and metastasis in cancer stem cells

    PubMed Central

    Liu, Chen-Chi; Lin, Shih-Pei; Hsu, Han-Shui; Yang, Shung-Haur; Lin, Chiu-Hua; Yang, Muh-Hwa; Hung, Mien-Chie; Hung, Shih-Chieh

    2016-01-01

    Targeting tumour-initiating cells (TICs) would lead to new therapies to cure cancer. We previously demonstrated that TICs have the capacity to survive under suspension conditions, while other cells undergo anoikis. Here we show that TICs exhibit increased phosphorylation levels of S727STAT3 because of PP2A inactivation. Collagen 17 gene expression is upregulated in a STAT3-dependent manner, which also stabilizes laminin 5 and engages cells to form hemidesmosome-like junctions in response. Blocking the PP2A-S727STAT3-collagen 17 pathway inhibits the suspension survival of TICs and their ability to form tumours in mice, while activation of the same pathway increases the suspension survival and tumour-initiation capacities of bulk cancer cells. The S727STAT3 phosphorylation levels correlate with collagen 17 expression in colon tumour samples, and correlate inversely with survival. Finally, this signalling axis enhances the ability of TIC to form tumours in mouse models of malignant lung cancer pleural effusion and spontaneous colon cancer metastasis. PMID:27306323

  3. Fumigant toxicity of essential oils from Achillea millefolium (asteraceae) and Prangos ferulacea (Apiaceae) against Sitophilus granarius and S. oryzea (col.: Curculionidae)

    NASA Astrophysics Data System (ADS)

    Şimşek, Şeyda; Pekbey, Gamze; Yaman, Cennet

    2016-04-01

    In the present study, experiments were conducted to investigate fumigant toxicity of the essential oil from Achillea millefolium (Asteraceae) and Prangos ferulacea (Apiaceae) plants for adult grain weevil (Sitophilus granarius) and rice weevil (S. oryzea) in vitro conditions. The essential oils were isolated with the water distillation method by Neo-Clevenger apparatus. During the study 10% (v/v) doses of oils in 20 cc of compressed rubber-capped glass tubes were used. After 24 hours mortality rates of the essential oils were compared. For S. granarius the toxicity of A. millefolium (98.85%) was observed to be more effective than P. ferulace (28.73%). Similarly for S. oryzea, A. millefolium (100 %) was found much more toxic than P. ferulace (9.82%). At the results of the study the essential oil of the A. millefolium has been determined as a promising insecticidal component against both pests.

  4. Characterization of Small ColE1-Like Plasmids Conferring Kanamycin Resistance in Salmonella enterica subsp. enterica serovars Typhimurium and Newport

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multi-antibiotic resistant (MR) Salmonella enterica serovars Typhimurium and Newport are an increasing concern in human and animal health. Many strains are known to carry antibiotic resistance determinants on multiple plasmids, yet detailed information is scarce. Three plasmids conferring kanamycin...

  5. ASCA Observations of the Dwarf Novae SS Cyg and VW Hyi And Observations of the Precessing Disk Cataclysmic Variable TV Col

    NASA Technical Reports Server (NTRS)

    Schlegel, E.; Swank, Jean (Technical Monitor)

    2001-01-01

    The observations for both SS Cyg and VW Hyi were to be scheduled as Targets of Opportunity jointly with other satellites. The VW Hyi observation was obtained jointly with EUVE during a superoutburst. The XTE data were initially processed, revealing no detection. However, the XTE team improved the instrumental background model and distributed it in July 1998. A further improvement was made in August 1999. The improved models allow a better background subtraction, thereby detecting previously un-detected sources.

  6. 40 CFR 721.2140 - Carbo-poly-cycli-col azo-alkyl-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. 721.2140 Section 721.2140 Protection of Environment...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. (a) Chemical substance and significant new uses...-mino-al-kyl-car-bo-mon-o-cyc-lic ester, halogen acid salt (PMN P-88-1682) is subject to reporting...

  7. 40 CFR 721.2140 - Carbo-poly-cycli-col azo-alkyl-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. 721.2140 Section 721.2140 Protection of Environment...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. (a) Chemical substance and significant new uses...-mino-al-kyl-car-bo-mon-o-cyc-lic ester, halogen acid salt (PMN P-88-1682) is subject to reporting...

  8. 40 CFR 721.2140 - Carbo-poly-cycli-col azo-alkyl-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. 721.2140 Section 721.2140 Protection of Environment...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. (a) Chemical substance and significant new uses...-mino-al-kyl-car-bo-mon-o-cyc-lic ester, halogen acid salt (PMN P-88-1682) is subject to reporting...

  9. 40 CFR 721.2140 - Carbo-poly-cycli-col azo-alkyl-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. 721.2140 Section 721.2140 Protection of Environment...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. (a) Chemical substance and significant new uses...-mino-al-kyl-car-bo-mon-o-cyc-lic ester, halogen acid salt (PMN P-88-1682) is subject to reporting...

  10. 40 CFR 721.2140 - Carbo-poly-cycli-col azo-alkyl-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. 721.2140 Section 721.2140 Protection of Environment...-aminoalkyl-carbo-mono-cyclic ester, halogen acid salt. (a) Chemical substance and significant new uses...-mino-al-kyl-car-bo-mon-o-cyc-lic ester, halogen acid salt (PMN P-88-1682) is subject to reporting...

  11. Characterization of genetically engineered mouse models carrying Col2a1-cre-induced deletions of Lrp5 and/or Lrp6

    PubMed Central

    Schumacher, Cassie A; Joiner, Danese M; Less, Kennen D; Drewry, Melissa Oosterhouse; Williams, Bart O

    2016-01-01

    Mice carrying Collagen2a1-cre-mediated deletions of Lrp5 and/or Lrp6 were created and characterized. Mice lacking either gene alone were viable and fertile with normal knee morphology. Mice in which both Lrp5 and Lrp6 were conditionally ablated via Collagen2a1-cre-mediated deletion displayed severe defects in skeletal development during embryogenesis. In addition, adult mice carrying Collagen2a1-cre-mediated deletions of Lrp5 and/or Lrp6 displayed low bone mass suggesting that the Collagen2a1-cre transgene was active in cells that subsequently differentiated into osteoblasts. In both embryonic skeletal development and establishment of adult bone mass, Lrp5 and Lrp6 carry out redundant functions. PMID:26962465

  12. Transcriptional analysis of the Escherichia coli ColV-Ia plasmid pS88 during growth in human serum and urine

    PubMed Central

    2012-01-01

    Background The sequenced O45:K1:H7 Escherichia coli meningitis strain S88 harbors a large virulence plasmid. To identify possible genetic determinants of pS88 virulence, we examined the transcriptomes of 88 plasmidic ORFs corresponding to known and putative virulence genes, and 35 ORFs of unknown function. Results Quantification of plasmidic transcripts was obtained by quantitative real-time reverse transcription of extracted RNA, normalized on three housekeeping genes. The transcriptome of E. coli strain S88 grown in human serum and urine ex vivo were compared to that obtained during growth in Luria Bertani broth, with and without iron depletion. We also analyzed the transcriptome of a pS88-like plasmid recovered from a neonate with urinary tract infection. The transcriptome obtained after ex vivo growth in serum and urine was very similar to those obtained in iron-depleted LB broth. Genes encoding iron acquisition systems were strongly upregulated. ShiF and ORF 123, two ORFs encoding protein with hypothetical function and physically linked to aerobactin and salmochelin loci, respectively, were also highly expressed in iron-depleted conditions and may correspond to ancillary iron acquisition genes. Four ORFs were induced ex vivo, independently of the iron concentration. Other putative virulence genes such as iss, etsC, ompTp and hlyF were not upregulated in any of the conditions studied. Transcriptome analysis of the pS88-like plasmid recovered in vivo showed a similar pattern of induction but at much higher levels. Conclusion We identify new pS88 genes potentially involved in the growth of E. coli meningitis strain S88 in human serum and urine. PMID:22720670

  13. Suspension survival mediated by PP2A-STAT3-Col XVII determines tumour initiation and metastasis in cancer stem cells.

    PubMed

    Liu, Chen-Chi; Lin, Shih-Pei; Hsu, Han-Shui; Yang, Shung-Haur; Lin, Chiu-Hua; Yang, Muh-Hwa; Hung, Mien-Chie; Hung, Shih-Chieh

    2016-01-01

    Targeting tumour-initiating cells (TICs) would lead to new therapies to cure cancer. We previously demonstrated that TICs have the capacity to survive under suspension conditions, while other cells undergo anoikis. Here we show that TICs exhibit increased phosphorylation levels of S727STAT3 because of PP2A inactivation. Collagen 17 gene expression is upregulated in a STAT3-dependent manner, which also stabilizes laminin 5 and engages cells to form hemidesmosome-like junctions in response. Blocking the PP2A-S727STAT3-collagen 17 pathway inhibits the suspension survival of TICs and their ability to form tumours in mice, while activation of the same pathway increases the suspension survival and tumour-initiation capacities of bulk cancer cells. The S727STAT3 phosphorylation levels correlate with collagen 17 expression in colon tumour samples, and correlate inversely with survival. Finally, this signalling axis enhances the ability of TIC to form tumours in mouse models of malignant lung cancer pleural effusion and spontaneous colon cancer metastasis. PMID:27306323

  14. Gender Effects on the Major Selection Process--A Five-Year Study: Implications for Marketing Business Programs of Small Private Col1eges to Women.

    ERIC Educational Resources Information Center

    Kolhede, Eric

    2001-01-01

    This 5-year study of undergraduates at a small western private college revealed similarities and differences between males and females in their expectations of business programs (e.g., women's greater desire for experiential learning), which point to product development and promotional strategies that can be targeted toward female students. (EV)

  15. "Their pineal glands aglow": Theosophical physiology in Ulysses.

    PubMed

    Morrisson, Mark S

    2008-01-01

    This article argues that Joyce's engagements with the Theosophy of the Dublin literary world amount to more than simple parody. In Ulysses, Joyce portrays Theosophy's efforts to offer an alternative understanding of physiology to that of the medical establishment as a form of boundary work, an adaptation of the discourse of modern medical research to fashion modern mysticism as a science. Ultimately, Joyce rejects Theosophical physiology and its evolutionary scientism because it provides an unsatisfactory rhetorical body, a failed attempt to renegotiate the boundaries between scientific materialism and spirituality in the awkward modernity of Dublin in 1904. PMID:20836274

  16. In vitro synergy of colistin combinations against extensively drug-resistant Acinetobacter baumannii producing OXA-23 carbapenemase.

    PubMed

    Wei, Wenjuan; Yang, Haifei; Liu, Yanyan; Ye, Ying; Li, Jiabin

    2016-06-01

    Fifty extensively drug-resistant Acinetobacter baumannii (XDRAB) were isolated from patients. The chequerboard microdilution method was used to determine the in vitro activities of five colistin (COL)-based combinations including COL+fosfomycin (FOS), COL+rifampicin (RIF), COL+imipenem (IMP), COL+sulbactam (SUP) and COL+levofloxacin (LVX). The synergistic activity was evaluated by the fractional inhibitory concentration index (FICI). According to our results, the combination of COL was synergistic with FOS, RIF, IMP, SUP and LVX with the ratios of 50, 72, 88, 92 and 64%, respectively. When combined with COL, the other five agents showed increased antimicrobial activities. In addition, two of the combinations, COL+RIF and COL+IMP, were more active than the combinations of COL+FOS, COL+SUP and COL+LVX. More importantly, these combination regimens could exert synergistic effects at the sub-minimum inhibitory concentration (MIC) levels against XDRAB strains. PMID:25978105

  17. 17 CFR 210.12-12B - Open option contracts written.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    .... Col. A Col. B Col. C Col. D Col. E Name of issuer 1,2 Number of contracts 3 Exercise price Expiration date Value. 4 1 Information as to put options shall be shown separately from information as to call options. 2 Options of an issuer where exercise prices or expiration dates differ shall be...

  18. 17 CFR 210.12-12B - Open option contracts written.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    .... Col. A Col. B Col. C Col. D Col. E Name of issuer 1,2 Number of contracts 3 Exercise price Expiration date Value. 4 1 Information as to put options shall be shown separately from information as to call options. 2 Options of an issuer where exercise prices or expiration dates differ shall be...

  19. "Where Are You Going, Where Have You Been?" and the Fantasies of the Unconscious.

    ERIC Educational Resources Information Center

    Cioe, Paul

    2003-01-01

    Compares Joyce Carol Oates's short story "Where Are You Going, Where Have You Been?" to the song lyrics of Bob Dylan. Explores the influence Dylan's music had on Oates. Discusses different ways to approach teaching this work. (SG)

  20. Translating Word-Play: French-English, English-French.

    ERIC Educational Resources Information Center

    Gordon, W. Terrence

    1986-01-01

    The linguistic complexity of humor is illustrated with examples of word play translated from French to English and English to French. Examples from the writings of James Joyce and Marcel Proust are highlighted. (CB)

  1. 78 FR 71583 - Proposed Collection; Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-29

    .... ACTION: Notice. SUMMARY: In compliance with Section 3506(c)(2)(A) of the Paperwork Reduction Act of 1995... Command (USAPHC), 5158 Blackhawk Road, ATTN: Joyce Woods, (MCHB- CS-CP), Aberdeen Proving Ground, MD...

  2. Fishing in the Holy Waters.

    ERIC Educational Resources Information Center

    Lipschultz, Geri

    1986-01-01

    Describes a teaching method that uses Molly Bloom's soliloquy from James Joyce's "Ulysses" to teach punctuation to freshman writing students. Argues that the assignment helps students discover their power as orderers. (EL)

  3. Skindeep Ulysses.

    PubMed

    Freedman, Ariela

    2008-01-01

    This essay is about Joyce as an epidermist and Joyce as a chronicler and cataloguer of the "skindeep" surfaces of Dublin in Ulysses. The book is crowded with skins: tanned skins, blushing skins, skins enhanced by makeup and creams, skins marked by race or religion, skins legible and visible, skins imagined and inaccessible and associated with both authenticity and disguise. Skin in Joyce becomes, in Steven Connor's terms, in The Book of Skin, "a place of minglings; a mingling of places," a space where medical, cultural, and aesthetic meanings jostle and intersect and are inscribed and projected on the surface that both expresses and conceals the subject. A skin-deep analysis of Ulysses can reveal to us the entanglement of surface and depth that characterizes Joyce's novel. PMID:20836270

  4. PTSD: A Search for "Active Ingredients."

    ERIC Educational Resources Information Center

    Huber, Charles H.

    1997-01-01

    Family counselors working with individuals suffering the effects of trauma are encouraged to consider the "active ingredients" found by Charles Figley and Joyce Carbonell at Florida State University and reported in the two articles reviewed. (Author/MKA)

  5. COMMENT: Comment on `Dirac theory in spacetime algebra'

    NASA Astrophysics Data System (ADS)

    Baylis, William E.

    2002-06-01

    In contrast to formulations of the Dirac theory by Hestenes and by the present author, the formulation recently presented by Joyce (Joyce W P 2001 J. Phys. A: Math. Gen. 34 1991-2005) is equivalent to the usual Dirac equation only in the case of vanishing mass. For nonzero mass, solutions to Joyce's equation can be solutions either of the Dirac equation in the Hestenes form or of the same equation with the sign of the mass reversed, and in general they are mixtures of the two possibilities. Because of this relationship, Joyce obtains twice as many linearly independent plane-wave solutions for a given momentum eigenvalue as exist in the conventional theory. A misconception about the symmetry of the Hestenes equation and the geometric significance of the algebraic spinors is also briefly discussed.

  6. Good vibrations: "sirens," soundscapes, and physiology.

    PubMed

    Plock, Vike Martina

    2008-01-01

    This article establishes Joyce's ongoing interest in psychoacoustics and illustrates how much he drew, in the writing of the "Sirens" episode, on nineteenth-century sound experiments that were developed by the German physician Hermann von Helmholtz. It argues that Joyce consciously referenced nineteenth-century sound theories to explore the link between the emotional and sensory experience of music and the physical and physiological components of sound perception. PMID:20836272

  7. Oliver St John Gogarty.

    PubMed

    Clarke, R W

    1997-01-01

    Oliver St John Gogarty--Otolaryngologist to fashionable Edwardian Dublin--was a distinguished poet and a Senator in the fledgling Irish Free State after its establishment in 1922. He numbered amongst his acquaintances the poet William Butler Yeats, the novelist James Joyce and a host of political and literary persona who helped to shape modern Ireland. He was satirised as 'stately plump Buck Mulligan' in Joyce's novel Ulysses. PMID:9292124

  8. Functional and evolutionary characterization of the CONSTANS gene family in short-day photoperiodic flowering in soybean.

    PubMed

    Wu, Faqiang; Price, Brian William; Haider, Waseem; Seufferheld, Gabriela; Nelson, Randall; Hanzawa, Yoshie

    2014-01-01

    CONSTANS (CO) plays a central role in photoperiodic flowering control of plants. However, much remains unknown about the function of the CO gene family in soybean and the molecular mechanisms underlying short-day photoperiodic flowering of soybean. We identified 26 CO homologs (GmCOLs) in the soybean genome, many of them previously unreported. Phylogenic analysis classified GmCOLs into three clades conserved among flowering plants. Two homeologous pairs in Clade I, GmCOL1a/GmCOL1b and GmCOL2a/GmCOL2b, showed the highest sequence similarity to Arabidopsis CO. The mRNA abundance of GmCOL1a and GmCOL1b exhibited a strong diurnal rhythm under flowering-inductive short days and peaked at dawn, which coincided with the rise of GmFT5a expression. In contrast, the mRNA abundance of GmCOL2a and GmCOL2b was extremely low. Our transgenic study demonstrated that GmCOL1a, GmCOL1b, GmCOL2a and GmCOL2b fully complemented the late flowering effect of the co-1 mutant in Arabidopsis. Together, these results indicate that GmCOL1a and GmCOL1b are potential inducers of flowering in soybean. Our data also indicate rapid regulatory divergence between GmCOL1a/GmCOL1b and GmCOL2a/GmCOL2b but conservation of their protein function. Dynamic evolution of GmCOL regulatory mechanisms may underlie the evolution of photoperiodic signaling in soybean. PMID:24465684

  9. A study of the clinical and radiological features in a cohort of 93 patients with a COL2A1 mutation causing spondyloepiphyseal dysplasia congenita or a related phenotype.

    PubMed

    Terhal, Paulien A; Nievelstein, Rutger Jan A J; Verver, Eva J J; Topsakal, Vedat; van Dommelen, Paula; Hoornaert, Kristien; Le Merrer, Martine; Zankl, Andreas; Simon, Marleen E H; Smithson, Sarah F; Marcelis, Carlo; Kerr, Bronwyn; Clayton-Smith, Jill; Kinning, Esther; Mansour, Sahar; Elmslie, Frances; Goodwin, Linda; van der Hout, Annemarie H; Veenstra-Knol, Hermine E; Herkert, Johanna C; Lund, Allan M; Hennekam, Raoul C M; Mégarbané, André; Lees, Melissa M; Wilson, Louise C; Male, Alison; Hurst, Jane; Alanay, Yasemin; Annerén, Göran; Betz, Regina C; Bongers, Ernie M H F; Cormier-Daire, Valerie; Dieux, Anne; David, Albert; Elting, Mariet W; van den Ende, Jenneke; Green, Andrew; van Hagen, Johanna M; Hertel, Niels Thomas; Holder-Espinasse, Muriel; den Hollander, Nicolette; Homfray, Tessa; Hove, Hanne D; Price, Susan; Raas-Rothschild, Annick; Rohrbach, Marianne; Schroeter, Barbara; Suri, Mohnish; Thompson, Elizabeth M; Tobias, Edward S; Toutain, Annick; Vreeburg, Maaike; Wakeling, Emma; Knoers, Nine V; Coucke, Paul; Mortier, Geert R

    2015-03-01

    Type 2 collagen disorders encompass a diverse group of skeletal dysplasias that are commonly associated with orthopedic, ocular, and hearing problems. However, the frequency of many clinical features has never been determined. We retrospectively investigated the clinical, radiological, and genotypic data in a group of 93 patients with molecularly confirmed SEDC or a related disorder. The majority of the patients (80/93) had short stature, with radiological features of SEDC (n = 64), others having SEMD (n = 5), Kniest dysplasia (n = 7), spondyloperipheral dysplasia (n = 2), or Torrance-like dysplasia (n = 2). The remaining 13 patients had normal stature with mild SED, Stickler-like syndrome or multiple epiphyseal dysplasia. Over 50% of the patients had undergone orthopedic surgery, usually for scoliosis, femoral osteotomy or hip replacement. Odontoid hypoplasia was present in 56% (95% CI 38-74) and a correlation between odontoid hypoplasia and short stature was observed. Atlanto-axial instability, was observed in 5 of the 18 patients (28%, 95% CI 10-54) in whom flexion-extension films of the cervical spine were available; however, it was rarely accompanied by myelopathy. Myopia was found in 45% (95% CI 35-56), and retinal detachment had occurred in 12% (95% CI 6-21; median age 14 years; youngest age 3.5 years). Thirty-two patients complained of hearing loss (37%, 95% CI 27-48) of whom 17 required hearing aids. The ophthalmological features and possibly also hearing loss are often relatively frequent and severe in patients with splicing mutations. Based on clinical findings, age at onset and genotype-phenotype correlations in this cohort, we propose guidelines for the management and follow-up in this group of disorders. PMID:25604898

  10. Substitution of Aspartate for glycine 1018 in the Type III procollagen (COL3AI) gene causes type IV Ehlers-Danlos Syndrome: The mutated allele is present in most blood leukocytes of the asymptomatic and mosaic mother

    SciTech Connect

    Kontusaari, S.; Tromp, G.; Kuivaniemi, H.; Prockop, D.J. ); Stolle, C. ); Pope, F.M.

    1992-09-01

    A proband with arterial ruptures and skin changes characteristic of the type IV variant of Ehlers-Danlos syndrome was found to have a single-base mutation in the type III procollagen gene, which converted the codon for glycine at amino position 1018 to a codon for aspartate. (Amino acid positions are numbered by the standard convention in which the first glycine of the triple-helical domain of an [alpha] chain is number 1. The numbers of positions in the [alpha]1(III) chains can be converted to positions in the human pro[alpha](III) chain by adding 167.). Nucleotide sequencing of overlapping PCR products in which the two alleles were distinguished demonstrated that the mutation of glycine 1018 was the only mutation that changed the primary structure of type III procollagen. The glycine substitution markedly decreased the amount of type III procollagen secreted into the medium by cultured skin fibroblasts from the proband. It is surprising that the same mutation was found in about 94% of the peripheral blood leukocytes from the proband's asymptomatic 72-year-old mother. Other tissues from the mother contained the mutated allele; it was present in 0%-100% of different samples of hair cells and in about 40% of cells from the oral epithelium. Therefore, the mother was a mosaic for the mutation. Since the mutated allele was present in cells derived from all three germ layers, the results indicated that the mutation arose by the late blastocyst stage of development. The results also indicate that assays of blood leukocytes do not always reveal mosaicism or predict phenotypic involvement of tissues, such as blood vessels, that are derived from the same embryonic cells as are leukocytes. 66 refs., 6 figs., 1 tab.

  11. Menstruation in Ulysses.

    PubMed

    Mullin, Katherine

    2008-01-01

    This article investigates James Joyce's fascination with a wide variety of medical texts, sexual folklores, religious beliefs, and persistent superstitions about menstruation. That fascination finds its way into Ulysses, which draws upon a number of intertexts to inform a curiosity about the female body most strikingly articulated by Bloom, Molly, and Gerty MacDowell. These intertexts are not simply imported into the novel but are dismantled and interrogated, as Joyce exposes, rather than endorses, clichés of essential femininity. PMID:20836273

  12. Preparation and characterization of an advanced collagen aggregate from porcine acellular dermal matrix.

    PubMed

    Liu, Xinhua; Dan, Nianhua; Dan, Weihua

    2016-07-01

    The objective of this study was to extract and characterize an advanced collagen aggregate (Ag-col) from porcine acellular dermal matrix (pADM). Based on histological examination, scanning electron microscopy (SEM) and atomic force microscope (AFM), Ag-col was composed of the D-periodic cross-striated collagen fibrils and thick collagen fiber bundles with uneven diameters and non-orientated arrangement. Fourier transform infrared (FTIR) spectra of pADM, Ag-col and Col were similar and revealed the presence of the triple helix. Circular dichroism (CD) analysis exhibited a slightly higher content of α-helix but inappreciably less amount of random coil structure in Ag-col compared to Col. Moreover, imino acid contents of pADM, Ag-col and Col were 222.43, 218.30 and 190.01 residues/1000 residues, respectively. From zeta potential analysis, a net charge of zero was found at pH 6.45 and 6.11 for Ag-col and Col, respectively. Differential scanning calorimetry (DSC) study suggested that the Td of Ag-col was 20°C higher than that of Col as expected, and dynamic mechanical analysis (DMA) indicated that Ag-col possessed a higher storage modulus but similar loss factor compared to Col. Therefore, the collagen aggregate from pADM could serve as a better alternative source of collagens for further applications in food and biological industries. PMID:27039117

  13. Gene expression analysis in patients with traumatic anterior shoulder instability suggests deregulation of collagen genes.

    PubMed

    Belangero, Paulo Santoro; Leal, Mariana Ferreira; Figueiredo, Eduardo Antônio; Cohen, Carina; Pochini, Alberto de Castro; Smith, Marília Cardoso; Andreoli, Carlos Vicente; Belangero, Sintia Iole; Ejnisman, Benno; Cohen, Moises

    2014-10-01

    Shoulder dislocation occurs in 1-2% of the population. Capsular deformation is a key factor in shoulder dislocation; however, little is known about capsule biology. We evaluated, for the first time in literature, the expression of COL1A1, COL1A2, COL3A1 and COL5A1 in the antero-inferior, antero-superior and posterior regions of the glenohumeral capsule of 31 patients with anterior shoulder instability and eight controls. The expression of collagen genes was evaluated by quantitative reverse transcription-PCR. The expression of COL1A1, COL3A1 and the ratio of COL1A1/COL1A2 were increased in all three portions of the capsule in patients compared to controls (p < 0.05). COL1A2 expression was upregulated in the antero-superior and posterior sites of the capsule of patients (p < 0.05). The ratio of COL1A2/COL3A1 expression was reduced in capsule antero-inferior and posterior sites of patients compared to controls (p < 0.05). In the capsule antero-inferior site of patients, the ratios of COL1A1/COL5A1, CO1A2/COL5A1 and COL3A1/COL5A1 expression were increased (p < 0.05). In patients, COL1A1/COL5A1 was also increased in the posterior site (p < 0.05). We found deregulated expression of collagen genes across the capsule of shoulder instability patients. These molecular alterations may lead to modifications of collagen fibril structure and impairment of the healing process, possibly with a role in capsular deformation. PMID:25042113

  14. Life sciences payload definition and integration study. Volume 1: Executive summary. [carry-on laboratory for Spacelab

    NASA Technical Reports Server (NTRS)

    1974-01-01

    The definition and integration tasks involved in the development of design concepts for a carry-on laboratory (COL), to be compatible with Spacelab operations, were divided into the following study areas: (1) identification of research and equipment requirements of the COL; (2) development of a number of conceptual layouts for COL based on the defined research of final conceptual designs; and (4) development of COL planning information for definition of COL/Spacelab interface data, cost data, and program cost schedules, including design drawings of a selected COL to permit fabrication of a functional breadboard.

  15. Preface

    NASA Astrophysics Data System (ADS)

    Wesch, Werner; Wolf, Gerhard K.

    2000-05-01

    cols="2">col1" colsep="0"/>col2" colsep="0"/>col1" nameend="col2">International CommitteeJ.C. Barbour, USAF.F. Komarov, BelarusG. Battaglin, ItalyE.A. Kotomin, LatviaM. Behar, BrazilG. Marletta, ItalyJ.P. Biersack, GermanyC.J. McHargue, USAL.T. Chadderton,AustraliaK. Roessler, GermanyA. Dunlop, FranceK.S.A. Song, CanadaG. Foti, ItalyP. Thevenard, FranceH.Hosono, JapanP.D. Townsend, UKN. Itoh, JapanW.J. Weber, USAR. Kelly, ItalyW.Wesch, GermanyI. Khubeis, JordanG.K. Wolf, Germanycol1" nameend="col2">col1" nameend="col2">Program CommitteeJ.P. Biersack, BerlinK. Roessler, JülichW. Bolse, StuttgartW.Skorupa, RossendorfK. Gärtner, JenaH. Stafast, JenaE. Glaser, JenaB. Weber, JenaS. Klaumünzer, BerlinW. Wesch, Jena

  16. A New Tool for Discourse Analysis: The Vocabulary-Management Profile.

    ERIC Educational Resources Information Center

    Youmans, Gilbert

    1991-01-01

    Proposes the Vocabulary-Management Profile, a tool for discourse analysis. The number of new words introduced in a moving interval of text 35 words long is counted and a curve created by plotting the number of new words in a successive interval at the midpoint of the interval. Analyses of text by George Orwell and James Joyce are presented. (JL)

  17. Macbeth: A Poetry Workshop on Stage at Shakespeare's Globe Theatre.

    ERIC Educational Resources Information Center

    Park, Keith

    2002-01-01

    The use of works by T. S. Eliot, James Joyce, and Shakespeare in interactive and experiential sessions with pupils with severe disabilities are described. A case study in assessment is provided by evaluating responses of one adolescent and showing how a Shakespeare workshop enabled her to become an active learner. (Contains references.) (Author/CR)

  18. Feminist Research Methods.

    ERIC Educational Resources Information Center

    Coyner, Sandra

    1993-01-01

    Reviews six works by Connie Miller, Comp., and Corinna Treitel (1991); Margrit Eichler (1987); Carol A. B. Warren (1988); Beth Hartung, Ed., and others (1988); Joyce McCarl Nielsen, Ed. (1990); and Liz Stanley, Ed. (1990). Works span diversity of current writing about feminist methodologies, offer guidelines for producing nonsexist or feminist…

  19. Content-Based Curriculum for High-Ability Learners.

    ERIC Educational Resources Information Center

    VanTassel-Baska, Joyce, Ed.; Little, Catherine A., Ed.

    The 14 chapters in this volume are intended to facilitate differentiated curriculum development for gifted students that is substantive, rigorous, and in keeping with disciplinary structures. The chapters are: (1) "Content-Based Curriculum for High Ability Learners: An Introduction" (Joyce VanTassel-Baska); (2) "Accelerating Learning Experiences…

  20. The Practice of Writing.

    ERIC Educational Resources Information Center

    Lodge, David

    With the constant theme of the mysterious process of creativity running through its essays, this book discusses the work of some much admired 20th-century writers--Graham Greene, James Joyce, D.H. Lawrence, Henry Green, Kingsley Amis, Vladimir Nabokov, and Anthony Burgess. The book addresses the situation of the contemporary novelist, both…

  1. Three Epiphanic Fragments: Education and the Essay in Memory

    ERIC Educational Resources Information Center

    Aldridge, David

    2014-01-01

    Pádraig Hogan has argued for a powerful conception of education as epiphany that is illuminated by the work of Heidegger and Joyce. But what are we to make of Stephen Dedalus' intention (pretension?) to "Remember your epiphanies"? Developing the phenomenological Erinnerungsversuch or "essay in memory" of David Farrell…

  2. Pennsylvania Adult and Continuing Education Research Conference Proceedings (Monroeville, Pennsylvania, October 8, 1994).

    ERIC Educational Resources Information Center

    Dean, Gary J., Ed.; Ferro, Trenton R., Ed.

    This document contains 5 invited papers, 11 refereed papers, and 3 symposium papers from an adult and continuing education research conference. The invited papers are: "Community and Adult Education: A Conceptual Framework for Theory and Practice" (Gary J. Dean); "Understanding the Control of Learning within Grassroots Initiatives" (Joyce S.…

  3. La musique du mot et du concept, ou certains problemes de traduction poetique (The Music of Word and Concept, or Certain Problems in Poetic Translation)

    ERIC Educational Resources Information Center

    Maisonneuve, Roland

    1978-01-01

    The whole universe enters the poet's being to be eventually transformed by him/her into musical language. It is this music that the translator must reproduce. Excerpts from the poems of Patmore, Auden, Donne, Joyce and Sloate illustrate the discussion. Translation of mystical and religious poetry is given special attention. (Text is in French.)…

  4. A Study of the Concurrent Validity of a Group Reasoning Test Built from Piaget's Tasks.

    ERIC Educational Resources Information Center

    Stefanich, Greg; And Others

    Researchers have attempted to develop paper and pencil Piagetian tests that yield results equivalent to manipulative Piagetian tasks and that can be administered in significantly less time. This study is an attempt to partially replicate and extend the findings of research by Paul Ankeny and Lyle Joyce who found significant positive correlations…

  5. Using Enquiry-Based Learning Methods to Teach "Finnegans Wake" to Undergraduates

    ERIC Educational Resources Information Center

    Manista, Frank C.; Gillespie, Michael Patrick

    2011-01-01

    Many readers dismiss James Joyce's final novel as impossible to wade through, with its multilingual puns, songs, jokes, portmanteau words, allusions, scientific references, myths and legends. Given the kinetic elements of any reading experience, features particularly evident in "Finnegans Wake", reading inevitably becomes synonymous with…

  6. Super Searchers Go to School: Sharing Online Strategies with K-12 Students, Teachers, and Librarians

    ERIC Educational Resources Information Center

    Valenza, Joyce Kasman

    2005-01-01

    Twelve prominent K-12 educators and educator librarians share their techniques and tips for helping students become effective, life-long information users. Through a series of skillful interviews, Joyce Kasman Valenza--techlife@school columnist for the "Philadelphia Inquirer" and herself a tech-savvy high school librarian--gets the experts to…

  7. Developing Verbal Talent: Ideas and Strategies for Teachers of Elementary and Middle School Students.

    ERIC Educational Resources Information Center

    VanTassel-Baska, Joyce, Ed.; Johnson, Dana T., Ed.; Boyce, Linda Neal, Ed.

    This book provides ideas and strategies for developing verbal talents in elementary and middle school students. Chapters include: (1) "The Process of Talent Development" (Joyce VanTassel-Baska); (2) "Talent Identification and Development in the Language Arts" (A. Harry Passow); (3) "Reading, Writing, and the Construction of Meaning" (Nancy Nelson…

  8. Convergent Validity of Group Tests of Cognitive Development.

    ERIC Educational Resources Information Center

    Stefanich, Greg P.; And Others

    1983-01-01

    Investigated convergent validity of individual Piaget/Inhelder clinical task interviews with Reasoning Test (Ankney and Joyce), Logical Reasoning Test (Burney), and Classroom Test of Formal Operations (Lawson). Indicates that correlations obtained are not sufficiently strong to warrant selection/categorization of individual students based on their…

  9. Dawn of a New Day. A WKCSC Presentation.

    ERIC Educational Resources Information Center

    Hartmann, Joyce; And Others

    This panel contains presentations by four members of the Western Kansas Community Services Consortium (WKCSC) who are involved in either teaching or administering college programs in Kansas in which part-time students are participating. Joyce Hartmann underscores the importance of part-time students as growing consumers of college programming in…

  10. Illinois Adult Education Bridges: Promising Practices. Transition Highlights. Issue 4

    ERIC Educational Resources Information Center

    Bragg, Debra; Oertle, Kathleen Marie; Kim, Sujung; Kirby, Catherine; Taylor, Jason; Harmon, Tim; Liss, Loralea

    2011-01-01

    To enhance state-level adult education and employment policy, in 2007 the Joyce Foundation began the Shifting Gears (SG) initiative to assist six states (Illinois, Indiana, Michigan, Minnesota, Ohio and Wisconsin) to integrate adult education, workforce development and postsecondary education policies and improve job opportunities for low-skilled…

  11. 75 FR 69158 - Quarterly Publication of Individuals, Who Have Chosen To Expatriate, as Required by Section 6039G

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-10

    ... Callaghan John Arthur Campeau Pamela Gail Cannon Ralph George Carlson Helma Maria Carstairs Emma Jane... Jason Chua Theodore Yuan-Shiun Chung Hyeon Joo Clark Lisa Michele Clarke Elizabeth Ann Coldren Patricia... Jerome P. Auger Judy Ann Auyang William C. Azhar Shariq Azrieli Stephanie Joyce Bachman Mary...

  12. Who Dat Say (We) "Too Depraved to Be Saved"?: Re-Membering Katrina/Haiti (and beyond): Critical Studyin' for Human Freedom

    ERIC Educational Resources Information Center

    King, Joyce E.

    2011-01-01

    In this essay, Joyce King attempts to interrupt the calculus of human (un)worthiness and to repair the collective cultural amnesia that are legacies of slavery and that make it easy--hegemonically and dysconsciously--for the public to accept myths and media reports, such as those about the depravity of survivors of Hurricane Katrina in New Orleans…

  13. Centres of excellence.

    PubMed

    Watson, J M

    1980-05-16

    The present Government may not be enthusiastic about health centres. But Dr Joyce M. Watson, of Glasgow University Department of General Practice and based at Woodside Health Centre in Glasgow, writes with enthusiasm of their advantages for the practice of medicine and the care of patients. PMID:10247174

  14. Everyone's Challenge. Proceedings of the Literacy Conference (Edmonton, Alberta, October 9-12, 1990).

    ERIC Educational Resources Information Center

    Konrad, Abram G., Ed.

    The following presentations are included in this report on conference proceedings: Conference Opening (Joyce Fairbairn); Greetings (John Gogo); Special Address (Ramon J. Hnatyshyn); "Paulo Freire on Adult Education," an interview and panel discussion (Carlos Torres et al.); Keynote Addresses (Jonathan Kozol, John Gogo, Peter Calamai, Frank Ogden);…

  15. Renewing Schools.

    ERIC Educational Resources Information Center

    McChesney, Jim

    1997-01-01

    This publication reviews works on educational reform that represent attempts to do more than merely respond in knee-jerk fashion to political pressure for reform. Bruce Joyce and Emily Calhoun, in "Learning Experiences in School Renewal: An Exploration of Five Successful Programs" (Eugene, Oregon: ERIC Clearinghouse on Educational Management,…

  16. Changing Course: Thurgood Marshall College Fund President Johnny Taylor Seeks New Partnerships and Avenues of Support for Public HBCUs

    ERIC Educational Resources Information Center

    Stuart, Reginald

    2011-01-01

    When veteran educator Dr. N. Joyce Payne handed the reins of the organization she founded, the Thurgood Marshall College Fund, to entertainment lawyer and board member Johnny Taylor, Taylor began pursuing a remake of the prestigious group that has turned it on its head in just a matter of months. Today, with just more than a year of leading the…

  17. Pedagogy Journal, 1994.

    ERIC Educational Resources Information Center

    Marashio Paul, Ed.; And Others

    1994-01-01

    This annual serial volume contains 20 articles offering practical pedagogical ideas from faculty at New Hampshire technical colleges. Section I, "Knowing a Thing," includes "A Rider Teaches Writing: Thoroughbreds and Freshmen," by Barbara Dimmick; "Some Thoughts on How To Incorporate Multimedia in Your Course," by Joyce Schneider; "Community…

  18. Proceedings of the Annual Meeting of the Association for Education in Journalism and Mass Communication (80th, Chicago, Illinois, July 30-August 2, 1997): Advertising.

    ERIC Educational Resources Information Center

    Association for Education in Journalism and Mass Communication.

    The Advertising section of the Proceedings contains the following 13 papers: "Offering a Creative Track in the Advertising Major: A Case History" (Beth E. Barnes and Carla V. Lloyd); "Messages of Individualism in French, Spanish, and American Television Advertising" (Ronald E. Taylor and Joyce Wolburg); "Frequency Levels and Activity Level…

  19. Moving from Inputs to Outputs to Outcomes: The Future of Education Policy

    ERIC Educational Resources Information Center

    Horn, Michael B.; Mackey, Katherine

    2011-01-01

    Over the previous two years, Innosight Institute has published a series of case studies about the fast-growing field of online learning in K-12 education. One aim of this effort was to identify policies that had either positive or negative ramifications for students. This policy brief, sponsored by the Joyce Foundation, summarizes some of that…

  20. An Online Peer Assisted Learning Community Model and its Application in ZJNU

    ERIC Educational Resources Information Center

    Gaofeng, Ruan; Yeyu, Lin

    2007-01-01

    Peer coaching, or peer assisting, was established in 1970s by Joyce and Showers. Initially used in teachers' professional development, it refers to a process that two or more teacher peers evaluate current practice mutually; expand skills, extract and build new skills; share ideas, and review & solve problems of classroom teaching in a way of…

  1. Realizing the Promise of 21st-Century Education: An Owner's Manual

    ERIC Educational Resources Information Center

    Joyce, Bruce; Calhoun, Emily

    2012-01-01

    While many futurists tout the value of teaching students 21st-century skills, bridging the concept with the practice is best accomplished by professional educators. Authors Bruce Joyce and Emily Calhoun know how to actualize the critical reforms that enable schools to prepare students for today's workforce. They outline a clear vision for…

  2. 78 FR 53755 - Good Neighbor Environmental Board; Notification of Public Advisory Committee Teleconference

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-30

    .... For further information regarding the teleconference and background materials, please contact Mark.../gneb . If you wish to make oral comments or submit written comments to the Board, please contact Mark... for individuals with disabilities, please contact Mark Joyce at (202) 564- 2130 or email at...

  3. Between the Cracks of History: Essays on Teaching and Illustrating Folklore. Publications of the Texas Folklore Society: 55.

    ERIC Educational Resources Information Center

    Abernethy, Francis Edward, Ed.; Satterwhite, Carolyn Fiedler, Ed.

    This book is composed of 21 essays that define and illustrate the folklore of Texas. Following the introduction, the six essays concerned with defining are: "Classroom Definitions of Folklore" (F. E. Abernethy); "Defining Folklore for My Students" (Joyce Roach); "Folklore and Cinema" (Jim Harris); "Toward a Definition of Folk Culture" (Joe S.…

  4. Economic Education Experiences of Enterprising Teachers. Volume 30.

    ERIC Educational Resources Information Center

    Nappi, Andrew T., Ed.

    This book describes award-winning teacher-developed projects and courses in economics. The reports are condensed versions of the original projects and are divided into grade levels. Primary Level includes: "Peanut Economics" (Janet Lancaster; Dena L. Meade); "Consumer Education Circus" (Pearl Eloshway; Linda McGeehan); "Critter Cards" (Joyce G.…

  5. Activities commemorating John B. Herrington as first Native American astronaut

    NASA Technical Reports Server (NTRS)

    2002-01-01

    KENNEDY SPACE CENTER, FLA. -- Chickasaw Nation Cultural Resources Director Haskell Alexander (left) presents a gift to Joyce and James Herrington, parents of John Herrington, mission specialist on mission STS-113. Herrington is the first Native American to be going into space.

  6. Anwendungsmoeglichkeiten der Textlinguistik im Englischunterricht der Sekundarstufe Zwei (Possibilities for the Use of Text Linguistics in English Teaching in Grades 11, 12, and 13).

    ERIC Educational Resources Information Center

    Mischkowski, Guenter

    1979-01-01

    Examines some findings from linguistically based text analyses for their possible utilization in teaching English in Grades 11-13 and in teaching literature. Notes some elements in text linguistics analogous to elements in present-day communication-oriented foreign language teaching, using James Joyce's short story "Eveline" for illustration.…

  7. Training a Non-Native Teacher.

    ERIC Educational Resources Information Center

    Ruwe, Donelle; Al-Wazedi, Umme

    2003-01-01

    Describes the differences between the second author's teaching experiences in Bangladesh and in the United States. Shares her approach to teaching James Joyce's "Araby," a short story commonly taught in high schools and in first-year undergraduate classes, and how her methods reflect differences in international approaches to education. Discusses…

  8. Becoming Literate: One Reader Reading.

    ERIC Educational Resources Information Center

    Wolf, Dennie Palmer

    1988-01-01

    This article presents an account of how one teacher and one student work through James Joyce's short story, "Eveline," illustrating an important reconceptualization of literacy--the teacher struggles to make public those aspects of reading which are often only assumed or left covert, namely, entering the world of the text, groping for meaning,…

  9. Activities commemorating John B. Herrington as first Native American astronaut

    NASA Technical Reports Server (NTRS)

    2002-01-01

    KENNEDY SPACE CENTER, FLA. -- Joyce and James Herrington, parents of John Herrington, accept a gift during a pre-launch Native American ceremony. They are the parents of John Herrington, mission specialist on mission STS-113. Herrington is the first Native American to be going into space.

  10. Woman as Reader: Pedagogical Implications.

    ERIC Educational Resources Information Center

    Flynn, Elizabeth A.

    Twenty-six women and 26 men enrolled in a humanities course at Michigan Technological University wrote their initial impressions to short stories by James Joyce, James Baldwin, Doris Lessing, and Virginia Woolf. The names were removed and the journal entries were analyzed in light of these four questions: (1) Do women refer to their personal…

  11. Differential Teaching of English Literature in Mixed Ability Classes.

    ERIC Educational Resources Information Center

    Kornblueth, Ilana; Shonberg, Sheila

    1985-01-01

    Elaborates on the existence of group differences within the student population, the relevance of the differences to English literature instruction, and ways to deal with the differences. Patterns of response and possible teaching strategies are described with reference to the story "Eveline" by James Joyce. (AMH)

  12. A Model of Cultural Leadership. The Achievements of Privately Funded Vouchers.

    ERIC Educational Resources Information Center

    Meyerson, Adam

    1999-01-01

    Describes the privately-funded voucher movement implemented by J. Patrick Rooney, James R. Leininger, Michael S. Joyce, John T. Walton, and Theodore J. Forstmann. An important feature of most of the programs is that parents have to pay partial tuition. Vouchers resources are focused on poor children in inner cities. (MMU)

  13. Power and Light without Electricity.

    ERIC Educational Resources Information Center

    Lang, Frederick K.

    One way to elicit good writing from students is to expose them to great literature and have them write about the attributes they share with a character, or about the experiences they have had that parallel those in a work of fiction, or about how a work of fiction compares with real "life." Using James Joyce's "Dubliners" as a primary source,…

  14. Teaching Must Be More Productive.

    ERIC Educational Resources Information Center

    Brandt, Ron

    1987-01-01

    Overviews a section in the same "Educational Leadership" issue on improving teaching productivity. Highlights Harold Stevenson's article comparing mathematics education in the U.S., China, and Japan and Bruce Joyce's summary and analysis of the research literature on effective instructional strategies. (MLH)

  15. Compilation of Action Research Papers in English Education.

    ERIC Educational Resources Information Center

    Sherman, Thomas F.; Lundquist, Margaret

    This action research compilation contains two research projects: "Increasing Student Appreciation of Poetry through the Use of Contemporary Music" by Paul G. Senjem and "Are Men and Women Created Equal? Gender in the Classroom" by Jennifer Joyce Plitzuweit. The researcher/author of the first paper states that his goal was to make exposure to…

  16. Bridge Programs in Illinois: Summaries, Outcomes, and Cross-Site Findings

    ERIC Educational Resources Information Center

    Bragg, D.; Harmon, T.; Kirby, C.; Kim, S.

    2010-01-01

    An increasing number of jobs in today's workforce require postsecondary education, yet large numbers of workers lack the essential skills and credentials to fill these jobs. The result is that many workers remain underemployed, reaching a ceiling early in their working careers. In 2007, the Joyce Foundation launched the Shifting Gears initiative…

  17. The State of the Art of Simulation in Teacher Education.

    ERIC Educational Resources Information Center

    Cruickshank, Donald R.; And Others

    1979-01-01

    Reviews the use of simulation in teacher preparation, with descriptions of the extent and the nature of such use. Data are taken from the reports of surveys conducted by Johnson, 1968; Sherwin, 1973; and Kuuskraa et al., Morra et al., and Joyce et al., 1977. Thirty-six references are listed. (Author/CMV)

  18. Self-Disclosure Decisions of University Students with Learning Disabilities

    ERIC Educational Resources Information Center

    Cole, Emma V.; Cawthon, Stephanie W.

    2015-01-01

    The number of students with learning disabilities (SLD) at postsecondary institutions has tripled over the past three decades and now constitutes about 11% of undergraduate students (Joyce & Rossen, 2006; U.S. Department of Education, 2013). Research has found that SLD who use accommodations at their postsecondary institution are more…

  19. Poetry at Buffalo: The Twentieth Century.

    ERIC Educational Resources Information Center

    Knoer, Wanda

    1981-01-01

    Discusses the Special Collections at the State University of New York at Buffalo and the role of Charles Abbott in their development. His interest and a serendipitous lack of funds have created an unusual collection containing a Twentieth Century Poetry Collection, James Joyce holographs, and rare book collections. (CHC)

  20. Planning Considerations for Afterschool Professional Development

    ERIC Educational Resources Information Center

    Bradshaw, L. Daniele

    2015-01-01

    Professional development is vital to the success of afterschool programs. Effective professional development enhances afterschool program quality by facilitating staff performance and knowledge; in addition, professional development is vital for improving student learning outcomes (Bouffard & Little, 2004; Hall & Surr, 2005; Joyce &…

  1. Observation Tools for Professional Development

    ERIC Educational Resources Information Center

    Malu, Kathleen F.

    2015-01-01

    Professional development of teachers, including English language teachers, empowers them to change in ways that improve teaching and learning (Gall and Acheson 2011; Murray 2010). In their seminal research on staff development--professional development in today's terms--Joyce and Showers (2002) identify key factors that promote teacher change.…

  2. Marketing Folk Art.

    ERIC Educational Resources Information Center

    Stevens, Phillips, Jr., Ed.

    1986-01-01

    "Marketing Folk Art" is a special section (pages 43-89) of this serial issue addressing the folklorists' role in developing marketing strategies to improve the lot of folk artists and protect their traditional forms of expression from commercial exploitation. The following six articles, introduced by Rosemary Joyce, focus on these topics: (1) "The…

  3. Instructional Models Effective in Distance Education.

    ERIC Educational Resources Information Center

    Jackman, Diane H.; Swan, Michael K.

    The purpose of this study was to identify which instructional models based on the framework of Joyce, Weil, and Showers, could be used effectively in distance education over the Interactive Video Network (IVN) system in North Dakota. Instructional models have been organized into families such as Information Processing, Social, Personal, and…

  4. A Teaching Repertoire

    ERIC Educational Resources Information Center

    Garrett, Joyce Lynn

    2007-01-01

    Building a repertoire of teaching skills means organizing instructional strategies in some meaningful way. In this brief essay, the author recommends that beginning teachers develop a repertoire comprised of a mix of strategies from each of Joyce, Weil, and Calhoun's (2004) categories: three strategies from the Cognitive Family (Concept…

  5. The Role of Aesthetic Theory in (Mass) Communications Theory.

    ERIC Educational Resources Information Center

    Theall, Donald F.

    This study in human communications ranges widely over scholarly work by Barthes, Bateson, Dewey, Richards, Peckham, Burke, and especially McLuhan, as well as art works by Warhol, Joyce, and various vodeo tape and film makers. In surveying the developments in art and advertising, the author finds that the arts (1) are now a kind of laboratory for…

  6. Professional Preparation in School Psychology: A Summary of Information from Programs in Seven Countries

    ERIC Educational Resources Information Center

    Oakland, Thomas; Hatzichristou, Chryse

    2014-01-01

    This article summarizes prominent themes found in descriptions of school psychology programs in Estonia (Kikas, 2014), Greece (Hatzichristou & Polychroni, 2014), Hong Kong (Lam, 2014), Romania (Negovan & Dinca, 2014), Sweden (Schad, 2014), United Kingdom (Wood, 2014), and United States (Joyce & Rossen, 2014). This paper summarizes…

  7. 78 FR 19725 - National Heart, Lung, and Blood Institute; Notice of Closed Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-02

    ... HUMAN SERVICES National Institutes of Health National Heart, Lung, and Blood Institute; Notice of Closed... of Committee: National Heart, Lung, and Blood Institute Special Emphasis Panel; Training in Lung... Joyce, Ph.D., Scientific Review Officer, Office of Scientific Review/DERA, National Heart, Lung,...

  8. Innovations in Labor Market Information and Their Application: Applications for Workforce Programs. A Greenways Action Brief

    ERIC Educational Resources Information Center

    Milfort, Myriam; Kelley, Jeremy

    2012-01-01

    With funding from the Joyce and Lumina foundations, Jobs for the Future (JFF) launched Credentials that Work to help postsecondary institutions, regions, and states align their occupational training programs to changing market demands. This initiative incorporates innovations in real-time labor market information in guiding institutions to better…

  9. Advances in Special Education Volume 11, Issues, Practices and Concerns in Special Education.

    ERIC Educational Resources Information Center

    Rotatori, Anthony F., Ed.; Schwenn, John O., Ed.; Burkhardt, Sandra, Ed.

    This volume presents 14 papers which address current issues and practices in special education. The papers are: (1) "National Educational Reform: General and Special Education" (Joyce Fiddler and Freddie W. Litton); (2) "Linguistically Appropriate Special Education" (Herbert Grossman); (3) "Portfolio Assessment: An Individualized Approach for…

  10. Poetic Voices: Writing, Reading, and Responding to Poetry

    ERIC Educational Resources Information Center

    Bandre, Patricia E.

    2012-01-01

    "Poetic Voices: Writing, Reading, and Responding to Poetry" was the title of the 2011 Master Class in Children's Literature. Woven into this session were the insights of poets Joyce Sidman and Pat Mora who shared their creative processes and the voices that inspire their poetry. In addition, Barbara Kiefer provided advice regarding how to connect…

  11. Tenured and Non-Tenured Teacher Perceptions on the Impact of District Designed Professional Development Courses on Classroom Practice

    ERIC Educational Resources Information Center

    Whitman, Joan Wrobleski

    2013-01-01

    Designers of professional development training often presume that teachers are able to apply new concepts classroom practice, but fail to include teacher voice, provide systemic follow-up, collegial support, and evaluation (Guskey, 2002; Joyce & Calhoun, 2010; McAdams, 2007). The study investigated differences between new, non-tenured and…

  12. 75 FR 9197 - Tennessee Gas Pipeline Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-01

    ... Energy Regulatory Commission Tennessee Gas Pipeline Company; Notice of Application February 19, 2010. Take notice that on February 9, 2010, Tennessee Gas Pipeline Company (Tennessee), 1001 Louisiana Street... application may be directed to Thomas G. Joyce, Manager, Certificates, Tennessee Gas Pipeline Company,...

  13. Literacy 2000: Make the Next Ten Years Matter. Conference Summary (New Westminster, British Columbia, October 18-21, 1990).

    ERIC Educational Resources Information Center

    Bossort, Patty, Ed.; And Others

    This conference summary contains 36 presentations. Participants' comments, taken from response cards, are quoted throughout. Presentations from the Opening Plenary include a keynote address--"What Is Literacy?: Critical Issues for the Next Decade" (John Ryan) and four "Panelist Presentations" (Francis Kazemek, Lorraine Fox, Joyce White, Robin…

  14. Activities commemorating John B. Herrington as first Native American astronaut

    NASA Technical Reports Server (NTRS)

    2002-01-01

    KENNEDY SPACE CENTER, FLA. - Chickasaw Dance troupe member Tim Harjo (second from left) leads Joyce and James Herrington in a dance honoring their son, STS-113 Mission Specialist John Herrington. The dance was part of a Native American ceremony at the Rocket Garden in the KSC Visitor Complex commemorating Herrington as the first tribally enrolled Native American astronaut to fly on a Shuttle mission.

  15. Deaf-Blind Perspectives, Fall 1994-Spring 1995.

    ERIC Educational Resources Information Center

    Deaf-Blind Perspectives, 1995

    1995-01-01

    This document consists of the three second-year issues of a newsletter concerning people with deaf-blindness. These issues include the following major articles: "A Report on Deaf-Blind Technical Assistance Collaboration" (Paddi Henderson and Rich Mulholland); "Rabbits and Retards" (Joyce Ford), in which a parent describes an encounter between her…

  16. The Targeted Reading Intervention: A Classroom Teacher Professional Development Program to Promote Effective Teaching for Struggling Readers in Kindergarten and First Grade

    ERIC Educational Resources Information Center

    Vernon-Feagans, Lynne; Kainz, Kirsten; Hedrick, Amy; Ginsberg, Marnie; Amendum, Steve

    2010-01-01

    The main objective of the overall Targeted Reading Intervention (TRI) was to help the classroom teacher acquire the key reading diagnostic strategies (e.g., Cooter, 2003; Desimone, 2009; Garet et al., 2001; Timperley & Phillips, 2003) relevant to K-1 struggling readers (e.g., Desimone, 2009; Garet et al., 2001; Guskey, 2002; Joyce & Showers, 2002;…

  17. A Computer Supported Simulator for Analyzing the Relationships Between Actual and Intended Teaching Behavior.

    ERIC Educational Resources Information Center

    Confessore, Gary J.

    A series of computer programs designed to provide a dynamic simulator of Joyce and Weil's models of interactive teaching are described. Initially developed and tested at the City University of New York (CUNY), the system is capable of supporting retrospective and predictive analysis of selected kinds and patterns of verbal behavior in a variety of…

  18. English Mini-Course World Literature (Preliminary, Unedited Version).

    ERIC Educational Resources Information Center

    Pittsburgh Board of Public Education, PA.

    This curriculum guide to a mini-course in world literature for high school students is restricted to the study of five short novels written in the twentieth century: "Demian" and "Siddhartha" by Hesse, "Platero and I" by Jimenez, "We Never Make Mistakes" by Solzhenitsyn, and "Portrait of the Artist as a Young Man" by Joyce. The objectives of the…

  19. 77 FR 23472 - Tennessee Gas Pipeline Company, L.L.C.; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-19

    ... Energy Regulatory Commission Tennessee Gas Pipeline Company, L.L.C.; Notice of Application Take notice that on April 4, 2012, Tennessee Gas Pipeline Company, L.L.C. (Tennessee), 1001 Louisiana Street... application may be directed to Thomas G. Joyce, Manager, Certificates, Tennessee Gas Pipeline Company,...

  20. 78 FR 68151 - Quarterly Publication of Individuals, Who Have Chosen To Expatriate, as Required by Section 6039G

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-13

    ... BAUREITHEL KARL HERBERT BEECHING SUSAN MARIE BENNETT SUSAN JANE BERAHA ALBERTO BERAHA BELLA CAROLINA BERAHA... KRASTEL-NICHOLS HELGA IRMA KREISELMEYER MARTHA MARIA KRUSELL PETER KARL KUBLIN JOYCE ARLENE KUSTER JANINE... TAN YANQIANG TANAKA RYO TANG MARX THERN EDINA RITTER THERN JR ROBERT WILLIAM TIEN LAUREN TSAK...