Blockade of alpha v beta3 and alpha v beta5 integrins by RGD mimetics induces anoikis and not integrin-mediated death in human endothelial cells.

Alpha v integrins are thought to play an important role in tumor angiogenesis. However, discrepancies between findings with Arg-Gly-Asp (RGD) mimetics, which block angiogenesis in animal models, and knockout mice, in which loss of some alpha v integrins enhances tumor angiogenesis, raise questions concerning the function of these integrins and the precise role of alpha v substrate mimetics in antiangiogenic therapies. We have examined the effects of a novel non-peptide RGD mimetic, S 36578-2, on human endothelial cells to elucidate its antagonist activity and to identify possible agonist functions.

Comparison of the NIST and BIPM Medium-Energy X-Ray Air-Kerma Measurements.

The air-kerma standards used for the measurement of medium-energy x rays were compared at the National Institute of Standards and Technology (NIST) and at the Bureau International des Poids et Mesures (BIPM). The comparison involved a series of measurements at the BIPM and the NIST using the air-kerma standards and two NIST reference-class transfer ionization standards. Reference beam qualities in the range from 60 kV to 300 kV were used.

A recurrent valpha17/vbeta10 TCR-expressing T cell clone is involved in the pathogenicity of collagen-induced arthritis in DBA/1 mice.

Collagen-induced arthritis (CIA) is an experimental model that mimics clinical and histological features of rheumatoid arthritis. In this disease, a crucial role in initiating the pathological changes has been assigned to T lymphocytes expressing the Th1 phenotype. Aiming at identifying type II collagen (CII)-specific T cells involved in CIA, T cell clones were generated in vitro from the lymph nodes (LN) of CII-immunized DBA / 1 mice.

Characterization of selachian egg case collagen.

The egg case of the dogfish Scyliorhinus canicula is a remarkable collagenous structure that combines mechanical strength and toughness with high permeability to small molecules and ions. The collagenous lamellae that form over 80% of the thickness of the case wall are secreted by the D-zone of the nidamental (oviducal gland). An acid-soluble collagen extracted from this zone and partially purified ran as a single band on a native gel at pH 4.

Volume recombination parameter in ionization chambers.

The parameter m2 governing the volume recombination in ionization chambers has been measured under conditions which allow strict application of the basic theory. The method consists of measuring the ratio of ionization currents I(V1) and I(V2) obtained at two given voltages V1 and V2 as a function of I(V1). The value of m2 is derived from a linear extrapolation to zero current.

Structural requirements for fibromodulin binding to collagen and the control of type I collagen fibrillogenesis--critical roles for disulphide bonding and the C-terminal region.

Fibromodulin belongs to the family of small, leucine-rich proteoglycans which have been reported to interact with collagens and to inhibit type I collagen fibrillogenesis. Decorin and fibromodulin exhibit a noticeable degree of sequence similarity. However, as previously reported [Font, B.

Processing of type XI collagen. Determination of the matrix forms of the alpha1(XI) chain.

Type XI collagen is mainly found as a minor constituent in type II-containing fibrils and presents a alpha1(XI)alpha2(XI)alpha3(XI) stoichiometry. This molecule was shown to be partially processed in its intact tissue form. Moreover, alternative splicing has been demonstrated in the variable region of the N-terminal domain of alpha1(XI) and alpha2(XI) chains.

Flexilin: a new extracellular matrix glycoprotein localized on collagen fibrils.

We have immunopurified and characterized a new glycoprotein of the extracellular matrix, using a monoclonal antibody obtained after immunization with fibril-associated collagens extracted from bovine tendon. In polyacrylamide gels, the protein migrates at about 350 kDa molecular mass. The protein is insensitive to bacterial collagenase, and no disulfide-linked aggregates could be detected; sugars were stained with periodic acid-Schiff's reagent.

Immunohistochemical and biochemical analyses of 20,000-25,000-year-old fossil cartilage.

A cracked, irregular pellicle adhering to fossilized bone excavated from the Enlène cave (Ariège) and estimated to date from 20,000-25,000 years BP was examined to verify its cartilaginous nature, suggested previously on the basis of optical and electron microscopic investigations. Immunolabeling of the organic component revealed the presence of type II and IX collagens, associated with residual glycosaminoglycans, in the external zone of the pellicle. The cartilaginous nature of the pellicle was also demonstrated by biochemical identification of type II collagen as the major protein in the demineralized sample: the amino acid compositions of insoluble and soluble fractions were similar to that of pure type II collagen; cyanogen-bromide-generated peptides, prepared after reduction of the sample, had an electrophoretic pattern similar to that of cyanogen bromide peptides derived from type II collagen.

Two-dimensional electrophoresis of intracellular and secreted protein synthesized by fetal bovine chondrocytes in high-density culture.

In order to study the mechanisms involved in the differentiation/dedifferentiation of chondrocytes, fetal bovine chondrocytes in high-density cultures were treated with retinoic acid, an agent known to modify the chondrocyte phenotype (10 mumol/L between day 2 to day 5 of culture). The synthesis of intracellular and secreted proteins was studied by two-dimensional electrophoresis in cell lysates and culture media after labeling with [35S]methionine for the last 14 h of culture. The proteins expressed in control and retinoic acid-treated cells were identified by microsequencing after "in-gel" tryptic digestion of the spot or by immunodetection with specific antibodies after two-dimensional gel blotting.

Onco-fetal/laminin-binding collagen from colon carcinoma: detection of new sequences.

We have recently identified an oncofetal-laminin binding collagen (OF/LB) composed of three alpha chains, with the apparent molecular mass of about 100 kDa each, but bearing different pI. One of the chains appears markedly acidic in a bidimensional electrophoretic system, where the NEPHGE is used as first dimension separating gel, while the two more basic chains have similar migration as alpha 1(III) and alpha 1(I) collagen chains, respectively. Sequence analyses have been performed on CNBr-peptides, derived from pepsinized triple helical molecules and on tryptic fragments obtained after in gel digestion of the acidic band.

Effect of retinoic acid on protein synthesis by foetal bovine chondrocytes in high-density culture: down-regulation of the glucose-regulated protein, GRP-78, and type II collagen.

The effect of 0.1-10 microM retinoic acid (RA) on foetal bovine chondrocytes was investigated in high-density cultures (0.6 x 10(6) cells/cm2).

Diversity in the processing events at the N-terminus of type-V collagen.

The processing of human collagen type-V chains was studied using anti-peptide polyclonal antibodies raised against peptide sequences at the N-terminal non-triple-helical region of pro-alpha 1(V) and pro-alpha 2(V) chains. The anti-peptide polyclonal antibody raised against positions 48-57 of the N-terminal alpha 2(V) sequence recognized the mature form of the human alpha 2(V) chain extracted without any proteolytic treatment from several tissues in the presence of a mixture of protease inhibitors. It also recognized the pro-alpha 2(V) and pN-alpha 2(V) collagen chains secreted in the cell-culture media of the rhabdomyosarcoma A204 cell line.

Characterization of type I procollagen N-proteinase from fetal bovine tendon and skin. Purification of the 500-kilodalton form of the enzyme from bovine tendon.

Procollagen N-proteinase (EC 3.4.24.

Lymphatic transport of cholesterol from exogenous and biliary origins in nonfasting rats after intraduodenal infusions of triolein.

Labelled thoracic-duct lymph was collected from nonfasting rats with a bile fistula after simultaneous intraduodenal infusions of bile labelled with [1-2 3H] cholesterol and a nutritive mixture containing [4-14C] cholesterol. The gastrointestinal tract, feces, chylomicrons and infranatants were analysed. Both biliary and exogenous cholesterol were absorbed by lymphatic way but the recovery of 3H labelling in total lymph was markedly higher than that of 14C activity.