Posttranslational control of membrane-skeleton (ankyrin and alpha beta- spectrin) assembly in early myogenesis

Adult chicken skeletal muscle cells express polypeptides that are antigenically related to alpha-spectrin (Mr 240,000) and beta-spectrin (Mr 220,000-225,000), the major components of the erythrocyte membrane- skeleton, and to ankyrin (Mr 237,000; also termed goblin in chicken erythrocytes), which binds spectrin to the transmembrane anion transporter in erythrocytes. Comparative immunoblotting of SDS- solubilized extracts of presumptive myoblasts and fully differentiated myotubes cultured in vitro demonstrated that there is a dramatic accumulation of ankyrin and alpha- and beta-spectrin during myogenesis and a concomitant switch in the subunit composition of spectrin from alpha gamma to alpha beta. Analysis of early time points in myogenesis (12-96 h) revealed that these changes occur shortly after the main burst of cell fusion. To determine the temporal relationship between cell fusion and the accumulation of ankyrin and alpha- and beta- spectrin, we treated presumptive myoblasts with 2 mM EGTA, which resulted in the complete inhibition of cell fusion. The incorporation of [35S]methionine into total protein and, specifically, into alpha-, gamma-, and beta-spectrin remained the same in EGTA-treated and control cells. Analysis by immunoblotting of the amounts of ankyrin and alpha- and beta-spectrin in fusion-blocked cells revealed that there was no effect on accumulation for the first 19 h. However, there was then a dramatic cessation in their accumulation, and thereafter, the amount of each protein at steady state remained constant. Upon release from the EGTA block, the cells fused rapidly (less than 11 h), and the accumulation of ankyrin and alpha- and beta-spectrin was reinitiated after a lag period of 3-5 h at a rate similar to that in control cells. The inhibition in the accumulation of newly synthesized ankyrin, alpha- spectrin, and beta-spectrin in EGTA-treated myoblasts was not characteristic of all structural proteins, since the accumulation of the muscle-specific intermediate filament protein desmin was the same in control and fusion-blocked cells. These results show that in myogenesis, the synthesis of ankyrin and alpha- and beta-spectrin and their accumulation as a complex, although concurrent, are not coupled events. We hypothesize that the extent of assembly of these components of the membrane-skeleton in muscle cells is determined by a control mechanism(s) operative at the posttranslational level that is triggered near the time of cell fusion and the onset of terminal differentiation

Pairwise interactive knowledge and Nash equilibrium

We provide epistemic conditions for Nash equilibrium, which are considerably weaker than thestandard ones by Aumann and Brandenburger (1995). Indeed, we simultaneously replace commonknowledge of conjectures and mutual knowledge of rationality with strictly weaker epistemicconditions of pairwise common knowledge of conjectures and pairwise mutual knowledge ofrationality respectively. It is also shown that, unlike the Aumann and Brandenburger''s conditions,ours do not imply common knowledge of rationality. Surprisingly, they actually do not even implymutual knowledge of rationality.microeconomics ;

Avian lens spectrin: subunit composition compared with erythrocyte and brain spectrin

Chicken lens spectrin is composed predominantly of equimolar amounts of two polypeptides with solubility properties similar, but not identical, to erythrocyte spectrin. The larger polypeptide, Mr 240,000 (lens alpha- spectrin), co-migrates with erythrocyte and brain alpha-spectrin on one- and two-dimensional SDS polyacrylamide gels and cross-reacts with antibodies specific for chicken erythrocyte alpha-spectrin; the smaller polypeptide, Mr 235,000 (lens gamma-spectrin), co-migrates with brain gamma-spectrin and does not cross-react with either the alpha-spectrin antibodies specific for chicken erythrocyte beta-spectrin. Minor amounts of polypeptides antigenically related to erythrocyte beta- spectrin with a greater electrophoretic mobility than lens gamma- spectrin are also detected in lens. The equimolar ratio of lens alpha- and gamma-spectrin is invariantly maintained during the extraction of lens plasma membranes under different conditions, or after immunoprecipitation of whole extracts of lens with erythrocyte alpha- spectrin antibodies. Two-dimensional peptide mapping reveals that whereas alpha-spectrins from chicken erythrocytes, brain, and lens are highly homologous, the gamma-spectrins, although related, have some cell-type-specific peptides and are substantially different from erythrocyte beta-spectrin. Thus, the expression of cell-type-specific gamma- and beta-spectrins may be the basis for the assembly of a spectrin-plasma membrane complex whose molecular composition is tailored to the functional requirements of the particular cell-type

A comparison of the distribution of actin and tubulin in the mammalian mitotic spindle as seen by indirect immunofluorescence

Rabbit antibodies against actin and tubulin were used in an indirect immunofluorescence study of the structure of the mitotic spindle of PtK1 cells after lysis under conditions that preserve anaphase chromosome movement. During early prophase there is no antiactin staining associated with the mitotic centers, but by late prophase, as the spindle is beginning to form, a small ball of actin antigenicity is found beside the nucleus; After nuclear envelope breakdown, the actiactin stains the region around each mitotic center, and becomes organized into fibers that run between the chromosomes and the poles. Colchicine blocks this organization, but does not disrupt the staining at the poles. At metaphase the antiactin reveals a halo of ill-defined radius around each spindle pole and fibers that run from the poles to the metaphase plate. Antitubulin shows astral rays, fibers running from chromosomes to poles, and some fibers that run across the metaphase plate. At anaphase, there is a shortening of the antiactin-stained fibers, leaving a zone which is essentially free of actin-staining fluorescence between the separating chromosomes. Antitubulin stains the region between chromosomes and poles, but also reveals substantial fibers running through the zone between separating chromosomes. Cells fixed during cytokinesis show actin in the region of the cleavage furrow, while antitubulin reveals the fibrous spindle remnant that runs between daughter cells. These results suggest that actin is a component of the mammalian mitotic spindle, that the distribution of actin differs from that of tubulin and that the distributions of these two fibrous proteins change in different ways during anaphase

Fracture Resistance of Compromised Endodontically Treated Teeth Restored with Bonded Post and Cores: An in Vitro Study

Statement of problem: It is unclear which post and core system performs best when bonded to severely compromised endodontically treated teeth. Purpose: The purpose of this study was to investigate the fracture resistance and mode of failure of severely compromised teeth restored with 3 different adhesively bonded post and core systems. Material and methods: Thirty extracted endodontically treated maxillary anterior teeth were randomly divided into 3 groups, CPC, gold cast post and core; TPC, titanium prefabricated post/composite resin core; and FPC, quartz fiber reinforced post/composite resin core. All posts were adhesively cemented. All cores resembled a central incisor preparation with no remaining tooth structure above the finish line. Cast gold crowns were fabricated and cemented adhesively. The specimens were aged with thermocycling and cyclic loading. Two specimens per group were randomly selected for micro-computed tomographic imaging before and after aging. Failure was induced with a universal testing machine. The mode of failure was characterized by the interface separation. Data were analyzed with 1-way ANOVA (α=.05) followed by post hoc tests (Bonferroni). Results: A statistically significant difference was found among the 3 groups (P=.002). CPC was significantly different than TPC (P=.008) or FPC (P=.003). The primary mode of failure for CPC and TPC was root fracture, and for FPC post debonding. Conclusions: Severely compromised endodontically treated teeth restored with bonded gold cast post and cores showed significantly higher fracture resistance

Source localization using acoustic vector sensors: a music approach

Traditionally, a large array of microphones is used to localize multiple far field sources in acoustics. We present a sound source localization technique that requires far less channels and measurement locations (affecting data channels, setup times and cabling issues). This is achieved by using an acoustic vector sensor (AVS) in air that consists of four collocated sensors: three orthogonally placed acoustic particle velocity sensors and an omnidirectional sound pressure transducer. Experimental evidence is presented demonstrating that a single 4 channel AVS based approach accurately localizes two uncorrelated sources. The method is extended to multiple AVS, increasing the number of sources that can be identified. Theory and measurement results are presented. Attention is paid to the theoretical possibilities and limitations of this approach, as well as the signal processing techniques based on the MUSIC method

The violent interstellar medium in Messier 31

Taurus observations in the line of H alpha and Very Large Array (VLA) HI mapping of the HII complex No. 722 in M31, reveal what seems to be a spherical cavity 330 pc in diameter blown out by a stellar association of over 20(exp 6) years old. Evidence of induced star formation which was initiated less than 5(exp 6) years ago is present in the form of bright HII emission and numerous O, B and Wolf-Rayet stars which are found within the shell surrounding the cavity. The energy necessary to create the HI shell is estimated to be about 5(exp 51) erg

`Mass without mass' from thin shells in Gauss-Bonnet gravity

Five tensor equations are obtained for a thin shell in Gauss-Bonnet gravity. There is the well known junction condition for the singular part of the stress tensor intrinsic to the shell, which we also prove to be well defined. There are also equations relating the geometry of the shell (jump and average of the extrinsic curvature as well as the intrinsic curvature) to the non-singular components of the bulk stress tensor on the sides of the thin shell. The equations are applied to spherically symmetric thin shells in vacuum. The shells are part of the vacuum, they carry no energy tensor. We classify these solutions of `thin shells of nothingness' in the pure Gauss-Bonnet theory. There are three types of solutions, with one, zero or two asymptotic regions respectively. The third kind of solution are wormholes. Although vacuum solutions, they have the appearance of mass in the asymptotic regions. It is striking that in this theory, exotic matter is not needed in order for wormholes to exist- they can exist even with no matter.Comment: 13 pages, RevTex, 8 figures. Version 2: includes discussion on the well-defined thin shell limit. Version 3: typos fixed, a reference added, accepted for publication in Phys. Rev.