Identification of 130 kDa cell surface LDL-binding protein from smooth muscle cells as a partially processed T-cadherin precursor

Atypical cell surface lipoprotein-binding proteins of 105 kDa and 130 kDa are present in membranes of vascular smooth muscle cells. We recently identified the 105 kDa protein from human aortic media as T-cadherin, an unusual glycosylphosphatidylinositol (GPI)-anchored member of the cadherin family of cell adhesion proteins. The goal of the present study was to determine the identity of 130 kDa lipoprotein-binding protein of smooth muscle cells. We applied different approaches that included protein sequencing of purified protein from human aortic media, the use of human T-cadherin peptide-specific antisera, and enzymatic treatment of cultured cells with trypsin and GPI-specific phospholipase C. Our results indicate that the 130 kDa protein is a partially processed form of T-cadherin which is attached to the membrane surface of smooth muscle cells via a GPI anchor and contains uncleaved N-terminal propeptide sequence. Our data disclose that, in contrast to classical cadherins, T-cadherin is expressed on the cell surface in both its precursor (130 kDa) and mature (105 kDa) forms.     

Modulation of gramicidin channel structure and function by the aliphatic "spacer" residues 10, 12, and 14 between the tryptophans

In the linear gramicidins, the four aromatic residues at positions 9, 11, 13, and 15 are well-known to be important for the structure and function of membrane-spanning gramicidin channels. To investigate whether the "spacer" residues between the tryptophans in gramicidin A (gA) are important for channel structure and function, D-Leu-10, -12. and -14 of gA were replaced by Ala, Val, or Ile. (For practical reasons, the Ile substitutions were introduced into the enantiomeric gramicidin A-, gA-.) Circular dichroism spectra of [D-Ala10,12,14]gA, [D-Val10,12,14]gA, or [Ile10,12,14]gA- incorporated into sodium dodecyl sulfate micelles or 1, 2-dimyristoyl-sn-glycero-3-phosphocholine vesicles differ from the spectrum of the native [D-Leu10,12,14]gA. All the analogue spectra display reduced ellipticity at both 218 and 235 nm, indicating the presence of double-stranded conformers with the Ala analogue spectra showing the largest departure from the native gA spectra. Size-exclusion chromatograms of the Val and Ile analogues show both monomer and dimer peaks, accompanied by peak broadening; the chromatograms for the Ala analogue show broad, overlapping peaks and suggest the presence of higher oligomers and/or (rapidly) interconverting conformations. All three analogues form membrane-spanning channels, with the channel-forming potency of the Ala analogue being much less than that of gA or the other analogues. In 1.0 M CsCl, the conductance of each analogue channel is approximately 25% less than that of [D-Leu10,12,14]gA channels. The lifetimes of the analogue channels also are less than of [D-Leu10,12, 14]gA channels, with the largest (8-fold) reduction being for [D-Ala10,12,14]gA channels. Hybrid channel experiments show that the beta6.3-helical backbone folding pattern is retained in the channel-forming subunits and that the substitutions primarily influence ion entry. Both the bulk and the stereochemistry of the aliphatic residues between the tryptophans of gA are important for channel structure and function.     

Forced convection in a channel partly occupied by a bidisperse porous medium: Asymmetric case

This paper presents an analytic investigation of forced convection in parallel-plate channel partly occupied by a bidisperse porous medium and partly by a fluid clear of solid material, the distribution being asymmetrical. The walls of the channel are subject to an uniform heat flux; the flow is assumed to be hydrodynamically and thermally fully developed. The layer of a bidisperse porous medium is attached to one of the channel walls; it is modeled utilizing a two-velocity two-temperature formulation using Darcy’s law. The Beavers–Joseph boundary condition is employed at the bidisperse porous medium/clear fluid interface. The dependences of the Nusselt number on a conductivity ratio, a velocity ratio, a volume fraction, internal heat exchange parameter, and the position of the porous-fluid interface are investigated. Both cases of symmetric and asymmetric heating are investigated, which is specified by the asymmetry heating parameter introduced here. For the case of asymmetric heating, a singular behavior of the Nusselt number is found and explained.     

The Cheng–Minkowycz problem for natural convection about a vertical plate embedded in a tridisperse porous medium

The classical Cheng–Minkowycz study of convection past a vertical plate embedded in a porous medium has been extended to the case of a tridisperse porous medium (TDPM). The boundary-layer analysis leads to expressions for the velocity and temperature fields in terms of two geometrical parameters, two inter-phase momentum transfer parameters, two thermal diffusivity ratios, two permeability ratios, two thermal conductivity ratios, and two inter-phase heat transfer parameters. For the leading edge region, and for an inner layer, a similarity solution is obtained. This involves the first eight parameters, each of which is a characteristic of the TDPM.     

On the molecular weight determination of a poly(aryl-ether-ether-ketone) (PEEK)

Attempts have been made to determine the weight average molecular weight ($\text{M}\text{?}{}_{\mathrm{<mtext>W</mtext>}}$), the radius of gyration ($\text{R}\text{?}{}_{\mathrm{<mtext>G</mtext>}}$) and the second virial coefficient (A₂) of five samples of poly(ether-ether-ketone) (PEEK) by light scattering (LS) in concentrated sulphuric acid. Account has been taken of the sulphonation of the polymer. Correlations between LS-molecular weights, melt viscosities and intrinsic viscosities in sulphuric acid or in a $\text{50}\text{50}$ phenol-trichlorobenzene mixture have been established. Gel permeation chromatography (g.p.c.) analyses at 115°C have been performed in this latter solvent and two calibration methods were compared.     

The use of natural interferon alpha conjugated to a monoclonal antibody anti mammary epithelial mucin (Mc5) for the treatment of human breast cancer xenografts

An immunoconjugate composed of natural interferon alpha (nIFN alpha) bound in a noncleavable fashion to a monoclonal antibody (MoAb) recognizing a breast epithelial membrane mucin (Mc5) was used to to treat xenografts of a human mammary carcinoma cell line (MCF-7) growing in nude mice. The immunoconjugate (nIFN alpha/Mc5) was administered as 20 intralesional (i.l.) injections to 1 of 2 xenografts in each animal. It was found that nIFN alpha/Mc5 produced a significant enhancement of the growth inhibitory actions of nIFN alpha on the injected tumors. Further enhancement was obtained when nIFN gamma or nIFN gamma together with Mc5 (at a dose 10 times larger than that present in nIFN alpha/Mc5) were added to the immunoconjugate. Biodistribution experiments showed that the uptake of 125I-nIFN alpha/Mc5 by the tumors was greater and its elimination slower than for 125I-nIFN alpha alone or conjugated to irrelevant mouse IgG1. In addition, the immunoconjugate up-regulated the antigenic expression of a breast epithelial membrane mucin by the carcinoma cells, an up-regulation which was not significantly different from that produced by nIFN alpha alone. The contralateral noninjected tumors exposed to systemic levels of the immunoconjugate showed an enhancement of antitumor effects, but to a lesser extent than the injected tumors. These findings suggest that the enhancement of the growth inhibitory action of the immunoconjugate was related to the specific binding of Mc5 which targeted the IFN to the carcinoma cells and impeded its elimination. It is likely that the targeting was favored by the IFN-mediated up-regulation of antigenic expression by the carcinoma cells, thereby producing a cascade of interrelated effects. The results of this study point out the feasibility and potential usefulness of IFN treatment by means of immunoconjugates as well as the worth of pursuing and improving this form of therapy.     

Mixed convection with viscous dissipation and pressure work in a lid-driven square enclosure

Buoyant laminar flow in a square lid-driven enclosure is analysed. The vertical sides are kept isothermal at different temperatures, while the horizontal sides are insulated. Assisting mixed convection flow due to uniform motion of the top side is considered. The governing balance equations are solved numerically by employing a Galerkin finite element method. The effects of viscous dissipation and pressure work are taken into account. In order to investigate the influence of these effects, the Nusselt number is evaluated with respect to the heat fluxes at both vertical sides, for different values of the Rayleigh number and of the Péclet number based on the lid velocity. Two sample fluids are considered: a gas and a highly viscous liquid. In the framework of the Oberbeck–Boussinesq approximation, a comparison is made between three different energy balance models: (A) enthalpy formulation (pressure work and viscous dissipation are included); (B) internal-energy formulation (viscous dissipation is included); (C) both pressure work and viscous dissipation are neglected. It is shown that, in the absence of a lid motion, the three models yield substantially the same predictions. On the other hand, when the forced flow induced by the lid motion becomes sufficiently large, the three models yield discrepant results, thus implying that pressure work and viscous dissipation are not negligible. Moreover, it is shown that, in this case, model (A) yields unphysical results, while model (B) leads to reasonable predictions.     

Chain length of the polylysine in receptor-targeted gene transfer complexes affects duration of reporter gene expression both in vitro and in vivo

Complexes composed of peptide ligand for the serpin enzyme complex receptor covalently coupled to poly-L-lysine condensed by charge interaction with plasmid DNA direct gene transfer into receptor bearing cells. We compared intensity and duration of reporter gene expression in vitro and in vivo from serpin-enzyme receptor-directed gene transfer complexes prepared with poly-L-lysine of different chain lengths. When substituted with linker and ligand to comparable extents, DNA complexes containing short chain poly-L-lysine were larger and gave higher peak expression but significantly shorter duration of expression than those containing long chain poly-L-lysine. Both peak expression and duration of expression exceeded that observed with Lipofectin. Neither naked DNA nor DNA complexed with unsubstituted polylysine was effective in gene transfer. For in vivo experiments, complexes containing optimal ligand and degree of substitution (based on in vitro data, peptide C105Y, 11 ligands/plasmid DNA molecule) were prepared with either short chain or long chain polylysine and a beta-galactosidase expression plasmid. Following injection into the tail veins of mice, longer chain complexes gave significantly higher expression of reporter gene in lung and spleen that lasted for a significantly longer period of time than the shorter chain complexes. The short chain poly-L-lysine-DNA complexes were larger in diameter, as assessed by electron microscopy or atomic force microscopy, and gave less protection against DNase digestion in vitro than longer chain complexes. Thus, for gene transfer complexes directed at the serpin enzyme complex receptor, longer chain poly-L-lysine gave a much longer duration of expression both in vitro and in vivo. We speculate that this may be due to protection against degradation afforded the plasmid DNA by the tighter compaction produced by long chain poly-L-lysine.     

The value of the Parenting Scale for measuring the discipline practices of parents of middle school children

The psychometric properties of the Parenting Scale (Arnold, O'Leary, Wolff, and Acker, 1993), a 30-item instrument originally developed to assess the discipline practices of parents of preschool children, were examined for parents of middle school students. Subjects were 298 parents of middle school student identified as at-risk for problem behavior. An exploratory factor analysis identified two factors labeled 'Overreactivity' and 'Laxness', closely resembling two of the factors found by Arnold et al., but each of these factors contained only six items. Confirmatory factor analyses, using data from the first two assessments, replicated this factor structure. The factors were significantly correlated with measures of parents' behavior, with scales from the child Behavior Checklist and Parent Daily Reports, and with the Beck Depression Inventory. The Laxness factor was less robust than the Overreactivity factor.