Nacre Evolution : A Proteomic Approach

AbstractFrom an evolutionary viewpoint, the molluscan nacre constitutes a fascinating object. This microstructure appeared early, in the Lower Cambrian period, about 530 million years ago, and since then, has been kept unchanged until today. Nacre is restricted to the conchiferan mollusks, where it occurs in t least three main classes, bivalves, gastropods and cephalopods. The aim of the present study is to investigate whether all nacres are built from the same “macromolecular tools”, proteins of the nacre matrix. To this end, we studied three new nacre models, the freshwater bivalve Unio pictorum, the cephalopod Nautilus macromphalus, and the gastropod Haliotis asinina, to which we applied a combined biochemical and proteomic characterization of their respective nacre matrices. The results of our approach, that can be defined as “shellomics” (proteomics applied to shell proteins) shed a new light on the macroevolution of nacre matrix proteins and on the in vitro design of nacre-like biomaterials.

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Isolation and Characterization of Human Epidermal Stem Cells

Clinical Science ◽

10.1042/cs0910141 ◽

1996 ◽

Vol 91 (2) ◽

pp. 141-146 ◽

Cited By ~ 24

Author(s):

P. H. Jones

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Matrix Proteins ◽

Stem Cell Markers ◽

Epidermal Stem Cells ◽

Cell Behaviour ◽

Isolation And Characterization ◽

Integrin Family

1. The keratinocytes in human epidermis are constantly turned over and replaced by a population of stem cells located in the basal epidermal layer. Until recently there were no markers allowing the isolation of viable epidermal stem cells. However, it has now been shown that epidermal stem cells can be isolated both in vitro and direct from the epidermis as they express high levels of functional β1 integrin family receptors for extracellular matrix proteins. 2. The evidence for integrins as stem cell markers and the insights that have been gained into stem cell behaviour are reviewed.

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Characterization of DNA sequences localized 11 to 17.5 kb upstream of the chicken embryonic pi-globin gene.

Acta Biochimica Polonica ◽

10.18388/abp.1999_4150 ◽

1999 ◽

Vol 46 (3) ◽

pp. 777-784

Author(s):

M Pietrowska ◽

M Rusin ◽

P Widłak ◽

S V Razin ◽

J Rzeszowska-Wolny

Keyword(s):

Dna Sequences ◽

Globin Gene ◽

Matrix Proteins ◽

Inverted Repeats ◽

Nuclear Matrix Proteins ◽

Alpha Globin ◽

Dna Fragment ◽

Globin Gene Domain

We have analyzed the DNA fragment localized about 11 to 17.5 kb upstream of the chicken alpha-globin gene domain (the fragment was designed as alpha-0). The nucleotide sequence of its 3.3 kb-long 5' part was established and interactions with nuclear matrix proteins were studied. The DNA region localized about 16 kb upstream of the embryonic pi-globin gene showed high affinity to nuclear matrices in vitro. Two palindromes and a cluster of inverted repeats were co-localized in the same region. The whole 6.6 kb alpha-0 fragment decreased the activity of linked CAT reporter gene when transfected into chicken erythroblastoid cells.

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A Proteomic Approach to the Characterization of the Endothelial Dysfunction in Uremia.

Blood ◽

10.1182/blood.v106.11.3955.3955 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3955-3955

Author(s):

Carla Carbo ◽

Maribel Diaz-Ricart ◽

Berta Fuste ◽

Aleix Cases ◽

Montserrat Carrascal ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Protein Identification ◽

Two Dimensional Gel Electrophoresis ◽

Cell Dysfunction ◽

Proteomic Approach ◽

High Incidence ◽

Vitro Model ◽

Identification Techniques

Abstract Deficient hemostasis coexists paradoxically with accelerated atherosclerosis in patients with cronic renal failure (CRF). The high incidence of aterothrombotic events in these patients has been associated with endothelial dysfunction. We have established an in vitro model of cell dysfunction by exposing endothelial cells (EC) to uremic media (Serradell y cols, Thromb Haemost86:1099,2001). However, it is difficult to advance in this research using conventional protein identification techniques. We have applied a proteomic approach to exhaustively characterize the EC dysfunction in uremia. Extracts of confluent EC, grown in the presence of sera from control donors and from patients under hemodialysis, were collected and lysated. Proteins were resolved by molecular weigh (Mr range 10–50kDa) and isoelectric point (pI range 3–10) using two-dimensional gel electrophoresis (2-DE). Spots were visualized by silver staining and further identified by mass spectrometry (MALDI-TOF). A search for significant changes in protein phosphorylation and glycosilation, and overexpression or new presence of proteins was specifically performed in uremic EC. Identification of some of the most prominent proteins revealed molecules related to inflammation (HMGB1, 25148/5.8, Mr/pI; aldose reductase, 35854/6.5; a proteasome component, 29192/5.7) and oxidative stress (superoxide dismutase, 24772/8.3; gluthatione peroxidase, 21899/6.2), both processes associated with CRF. In addition, we observed a significant increase in the protein destrin (18506/8.1), an actin depolymerizing protein. This finding could be related to the previously reported increased detachment of uremic EC when exposed to flowing blood. Therefore, proteomics can be used to compare the proteome of control and uremic EC, helping to reveal information on the molecular basis of the disease. A more exhaustive analysis of 2-DE gels will provide answers and potential therapeutic targets in a short future.

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In vitro characterization of neural stem cells: Functional response to the enteric neurotrophin GDNF and co-culture with human colonic smooth muscle

Gastroenterology ◽

10.1016/s0016-5085(01)80307-3 ◽

2001 ◽

Vol 120 (5) ◽

pp. A62-A62

Author(s):

M MICCI ◽

R LEARISH ◽

P PASRICHA

Keyword(s):

Stem Cells ◽

Smooth Muscle ◽

Neural Stem Cells ◽

Functional Response ◽

In Vitro Characterization

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Biological and physico-chemical characterization of ultra high diluted Hydrastis canadensis and in vitro studies on mammalian cells

Allgemeine Homöopathische Zeitung ◽

10.1055/s-0037-1601241 ◽

2017 ◽

Vol 262 (02) ◽

pp. 2-76

Author(s):

N Chandrakar ◽

MK Temgire ◽

SG Kane ◽

AK Suresh ◽

J Bellare

Keyword(s):

Mammalian Cells ◽

Chemical Characterization ◽

In Vitro Studies ◽

Hydrastis Canadensis ◽

Physico Chemical

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Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

Veterinary and Comparative Orthopaedics and Traumatology ◽

10.1055/s-0038-1632561 ◽

1997 ◽

Vol 10 (01) ◽

pp. 6-11 ◽

Cited By ~ 3

Author(s):

R. F. Rosenbusch ◽

L. C. Booth ◽

L. A. Dahlgren

Keyword(s):

Electron Microscopy ◽

Extracellular Matrix ◽

Light Microscopy ◽

Light And Electron Microscopy ◽

Tendon Healing ◽

Matrix Proteins ◽

Isolated Cells ◽

Superficial Digital Flexor Tendon ◽

Vitro Model

SummaryEquine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

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Assays for Phospholipid-Dependent Formation of Thrombin and Xa: A Potential Method for Quantifying Lupus Anticoagulant Activity

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646437 ◽

1991 ◽

Vol 66 (04) ◽

pp. 453-458 ◽

Cited By ~ 15

Author(s):

John T Brandt

Keyword(s):

Specific Activity ◽

Anticoagulant Activity ◽

Factor Xa ◽

Clinical Importance ◽

Potential Method ◽

Quantitative Expression ◽

Increased Risk ◽

Apparent Association

SummaryLupus anticoagulants (LAs) are antibodies which interfere with phospholipid-dependent procoagulant reactions. Their clinical importance is due to their apparent association with an increased risk of thrombo-embolic disease. To date there have been few assays for quantifying the specific activity of these antibodies in vitro and this has hampered attempts to purify and characterize these antibodies. Methods for determining phospholipid-dependent generation of thrombin and factor Xa are described. Isolated IgG fractions from 7 of 9 patients with LAs were found to reproducibly inhibit enzyme generation in these assay systems, permitting quantitative expression of inhibitor activity. Different patterns of inhibitory activity, based on the relative inhibition of thrombin and factor Xa generation, were found, further substantiating the known heterogeneity of these antibodies. These systems may prove helpful in further purification and characterization of LAs.

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Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648381 ◽

1992 ◽

Vol 67 (01) ◽

pp. 063-065 ◽

Cited By ~ 6

Author(s):

Sherryl A M Taylor ◽

Jacalyn Duffin ◽

Cherie Cameron ◽

Jerome Teitel ◽

Bernadette Garvey ◽

...

Keyword(s):

Nucleotide Substitution ◽

Novel Mutation ◽

Factor Ix ◽

Causative Mutation ◽

Coding Region ◽

Body Of Knowledge ◽

Polymerase Chain ◽

Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

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