Generation of specific antibodies against the rap1A, rap1B and rap2 small GTP-binding proteins. Analysis of rap and ras proteins in membranes from mammalian cells

Specific antibodies against rap1A and rap1B small GTP-binding proteins were generated by immunization of rabbits with peptides derived from the C-terminus of the processed proteins. Immunoblot analysis of membranes from several mammalian cell lines and human thrombocytes with affinity-purified antibodies against rap1A or rap1B demonstrated the presence of multiple immunoreactive proteins in the 22-23 kDa range, although at strongly varying levels. Whereas both proteins were present in substantial amounts in membranes from myelocytic HL-60, K-562 and HEL cells, they were hardly detectable in membranes from lymphoma U-937 and S49.1 cyc- cells. Membranes from human thrombocytes and 3T3-Swiss Albino fibroblasts showed strong rap1B immunoreactivity, whereas rap1A protein was present in much lower amounts. In the cytosol of HL-60 cells, only small amounts of rap1A and rap1B proteins were detected, unless the cells were treated with lovastatin, an inhibitor of hydroxymethylglutaryl-coenzyme A reductase, suggesting that both proteins are isoprenylated. By comparison with recombinant proteins, the ratio of rap1A/ras proteins in membranes from HL-60 cells was estimated to be about 4:1. An antiserum directed against the C-terminus of rap2 reacted strongly with recombinant rap2, but not with membranes from tested mammalian cells. In conclusion, rap1A and rap1B proteins are distributed differentially among membranes from various mammalian cell types and are isoprenylated in HL-60 cells

The 54-kD Protein of Signal Recognition Particle Contains a Methionine-rich RNA Binding Domain

Signal recognition particle (SRP) plays the key role in targeting secretory proteins to the membrane of the endoplasmic reticulum (Walter, P., and V. R. Lingappa. 1986. Annu. Rev. CeliBiol. 2:499-516). It consists of SRP7S RNA and six proteins. The 54-kD protein of SRP (SRP54) recognizes the signal sequence of nascent polypeptides. The 19-kD protein of SRP (SRP19) binds to SRP7S RNA directly and is required for the binding of SRP54 to the particle. We used deletion mutants of SRP19 and SRP54 and an in vitro assembly assay in the presence of SRP7S RNA to define the regions in both proteins which are required to form a ribonucleoprotein particle. Deletion of the 21 COOH-terminal amino acids of SRP19 does not interfere with its binding to SRP7S RNA. Further deletions abolish SRP19 binding to SRP7S RNA. The COOH-terminal 207 amino acids of SRP54 (M domain) were found to be necessary and sufficient for binding to the SRP19/7S RNA complex in vitro. Limited protease digestion of purified SRP confirmed our results for SRP54 from the in vitro binding assay. The SRP54M domain could also bind to Escherichia coli 4.5S RNA that is homologous to part of SRP7S RNA. We suggest that the methioninerich COOH terminus of SRP54 is a RNA binding domain and that SRP19 serves to establish a binding site for SRP54 on the SRP7S RNA

An approximate model for cancellous bone screw fixation

This is the author's accepted manuscript. The final published article is available from the link below. Copyright @ 2013 Taylor & Francis.This paper presents a finite element (FE) model to identify parameters that affect the performance of an improved cancellous bone screw fixation technique, and hence potentially improve fracture treatment. In cancellous bone of low apparent density, it can be difficult to achieve adequate screw fixation and hence provide stable fracture fixation that enables bone healing. Data from predictive FE models indicate that cements can have a significant potential to improve screw holding power in cancellous bone. These FE models are used to demonstrate the key parameters that determine pull-out strength in a variety of screw, bone and cement set-ups, and to compare the effectiveness of different configurations. The paper concludes that significant advantages, up to an order of magnitude, in screw pull-out strength in cancellous bone might be gained by the appropriate use of a currently approved calcium phosphate cement

In-Line-Test of Variability and Bit-Error-Rate of HfOx-Based Resistive Memory

Spatial and temporal variability of HfOx-based resistive random access memory (RRAM) are investigated for manufacturing and product designs. Manufacturing variability is characterized at different levels including lots, wafers, and chips. Bit-error-rate (BER) is proposed as a holistic parameter for the write cycle resistance statistics. Using the electrical in-line-test cycle data, a method is developed to derive BERs as functions of the design margin, to provide guidance for technology evaluation and product design. The proposed BER calculation can also be used in the off-line bench test and build-in-self-test (BIST) for adaptive error correction and for the other types of random access memories.Comment: 4 pages. Memory Workshop (IMW), 2015 IEEE Internationa

Phonon anomalies and electron-phonon interaction in RuSr_2GdCu_2O_8 ferromagnetic superconductor: Evidence from infrared conductivity

Critical behavior of the infrared reflectivity of RuSr_2GdCu_2O_8 ceramics is observed near the superconducting T_{SC} = 45 K and magnetic T_M = 133 K transition temperatures. The optical conductivity reveals the typical features of the c-axis optical conductivity of strongly underdoped multilayer superconducting cuprates. The transformation of the Cu-O bending mode at 288 cm^{-1} to a broad absorption peak at the temperatures between T^* = 90 K and T_{SC} is clearly observed, and is accompanied by the suppression of spectral weight at low frequencies. The correlated shifts to lower frequencies of the Ru-related phonon mode at 190 cm^{-1} and the mid-IR band at 4800 cm^{-1} on decreasing temperature below T_M are observed. It provides experimental evidence in favor of strong electron-phonon coupling of the charge carriers in the Ru-O layers which critically depends on the Ru core spin alignment. The underdoped character of the superconductor is explained by strong hole depletion of the CuO_2 planes caused by the charge carrier self-trapping at the Ru moments.Comment: 11 pages incl. 5 figures, submitted to PR

The aza-Morita-Baylis-Hillman reaction of electronically and sterically deactivated substrates.

The aza-Morita–Baylis–Hillman (azaMBH) reaction has been studied for electronically and sterically deactivated Michael acceptors. It is found that electronically deactivated systems can be converted with electron-rich phosphanes and pyridines as catalysts equally well. For sterically deactivated systems clearly better catalytic turnover can be achieved with pyridine catalysts. This is in accordance with the calculated affinities of the catalysts towards different Michael-acceptors

The “floating forearm” injury in a child: a case report

The case of a eleven-year-old girl who had a fracture dislocation of the left elbow with entrapment of the ulnar nerve into the dislocated ulnar epicondyle anlage and unstable forearm fracture of the ipslateral upper extremity is described. This severe injury to the elbow and the ipsilateral forearm is termed “floating forearm” injury. The forearm was stabilized percutaneously and the elbow fracture dislocation, remaining unstable after internal fixation was treated with a pediatric elbow fixator with motion capacity

Variability of the pullout strength of cancellous bone screws with cement augmentation

Background Orthopaedic surgeons often face clinical situations where improved screw holding power in cancellous bone is needed. Injectable calcium phosphate cements are one option to enhance fixation. Methods Paired screw pullout tests were undertaken in which human cadaver bone was augmented with calcium phosphate cement. A finite element model was used to investigate sensitivity to screw positional placement. Findings Statistical analysis of the data concluded that the pullout strength was generally increased by cement augmentation in the in vitro human cadaver tests. However, when comparing the individual paired samples there were surprising results with lower strength than anticipated after augmentation, in apparent contradiction to the generally expected conclusion. Investigation using the finite element model showed that these strength reductions could be accounted for by small screw positional changes. A change of 0.5 mm might result in predicted pullout force changes of up to 28%. Interpretation Small changes in screw position might lead to significant changes in pullout strength sufficient to explain the lower than expected individual pullout values in augmented cancellous bone. Consequently whilst the addition of cement at a position of low strength would increase the pullout strength at that point, it might not reach the pullout strength of the un-augmented paired test site. However, the overall effect of cement augmentation produces a significant improvement at whatever point in the bone the screw is placed. The use of polymeric bone-substitute materials for tests may not reveal the natural variation encountered in tests using real bone structures.Dr V. Stadelmann (AOR, Davos, Switzerland) and Mr. M. Behrens (Stryker, Selzach, Switzerland). Professor Procter and Dr Arnoldi were employed by Stryker Trauma. Dr Bennani's PhD studies at Brunel University were funded by Stryker Trauma AG

Decoupled CuO_2 and RuO_2 layers in superconducting and magnetically ordered RuSr_2GdCu_2O_8

Comprehensive measurements of dc and ac susceptibility, dc resistance, magnetoresistance, Hall resistivity, and microwave absorption and dispersion in fields up to 8 T have been carried out on RuSr_2GdCu_2O_8 with the aim to establish the properties of RuO_2 and CuO_2 planes. At ~130 K, where the magnetic order develops in the RuO_2 planes, one observes a change in the slope of dc resistance, change in the sign of magnetoresistance, and the appearance of an extraordinary Hall effect. These features indicate that the RuO_2 planes are conducting. A detailed analysis of the ac susceptibility and microwave data on both, ceramic and powder samples show that the penetration depth remains frequency dependent and larger than the London penetration depth even at low temperatures. We conclude that the conductivity in the RuO_2 planes remains normal even when superconducting order is developed in the CuO_2 planes below \~45 K. Thus, experimental evidence is provided in support of theoretical models which base the coexistence of superconductivity and magnetic order on decoupled CuO_2 and RuO_2 planes.Comment: 11 pages, 11 figures, submitted to PR

Wind tunnel technology roadmap and analysis of the innovation within the field

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