The structure of the acto-myosin subfragment 1 complex: results of searches using data from electron microscopy and x-ray crystallography

Surmises of how myosin subfragment 1 (S1) interacts with actin filaments in muscle contraction rest upon knowing the relative arrangement of the two proteins. Although there exist crystallographic structures for both S1 and actin, as well as electron microscopy data for the acto-S1 complex (AS1), modeling of this arrangement has so far only been done "by eye." Here we report fitted AS1 structures obtained using a quantitative method that is both more objective and makes more complete use of the data. Using undistorted crystallographic results, the best-fit AS1 structure shows significant differences from that obtained by visual fitting. The best fit is produced using the F-actin model of Holmes et al. [Holmes, K. C., Popp, D., Gebhard, W. & Kabsch, W. (1990) Nature (London) 347, 44-49]. S1 residues at the AS1 interface are now found at a higher radius as well as being translated axially and rotated azimuthally. Fits using S1 plus loops missing from the crystal structure were achieved using a homology search method to predict loop structures. These improved fits favor an arrangement in which the loop at the 50- to 20-kDa domain junction of S1 is located near the N terminus of actin. Rigid-body movements of the lower 50-kDa domain, which further improve the fit, produce closure of the large 50-kDa domain cleft and bring conserved residues in the lower 50-kDa domain into an apparently appropriate orientation for close interaction with actin. This finding supports the idea that binding of ATP to AS1 at the end of the ATPase cycle disrupts the actin binding site by changing the conformation of the 50-kDa cleft of S1.     

Ancient mtDNA sequences in the human nuclear genome: a potential source of errors in identifying pathogenic mutations

Nuclear-localized mtDNA pseudogenes might explain a recent report describing a heteroplasmic mtDNA molecule containing five linked missense mutations dispersed over the contiguous mtDNA CO1 and CO2 genes in Alzheimer's disease (AD) patients. To test this hypothesis, we have used the PCR primers utilized in the original report to amplify CO1 and CO2 sequences from two independent rho degrees (mtDNA-less) cell lines. CO1 and CO2 sequences amplified from both of the rho degrees cells, demonstrating that these sequences are also present in the human nuclear DNA. The nuclear pseudogene CO1 and CO2 sequences were then tested for each of the five "AD" missense mutations by restriction endonuclease site variant assays. All five mutations were found in the nuclear CO1 and CO2 PCR products from rho degrees cells, but none were found in the PCR products obtained from cells with normal mtDNA. Moreover, when the overlapping nuclear CO1 and CO2 PCR products were cloned and sequenced, all five missense mutations were found, as well as a linked synonymous mutation. Unlike the findings in the original report, an additional 32 base substitutions were found, including two in adjacent tRNAs and a two base pair deletion in the CO2 gene. Phylogenetic analysis of the nuclear CO1 and CO2 sequences revealed that they diverged from modern human mtDNAs early in hominid evolution about 770,000 years before present. These data would be consistent with the interpretation that the missense mutations proposed to cause AD may be the product of ancient mtDNA variants preserved as nuclear pseudogenes.     

An effective back‐off selection technique for reliable beacon reception in VANETs

In safety‐critical scenarios, reliable reception of beacons transmitted by a subject vehicle is critical to avoid vehicle collision. According to the employed contention window sizes in IEEE 802.11p, beacons are transmitted with a small contention window size. As a result, multiple vehicles contend for the shared channel access by selecting the same back‐off slot. This is a perfect recipe for synchronous collisions wherein reliable beacon delivery cannot be guaranteed for any vehicle. We consider the problem of selecting the back‐off slots from the current contention window to provide reliable delivery of beacons transmitted by a subject vehicle to its neighbors. Given a safety scenario, we propose a Pseudo‐Random Number Generator (PRNG)‐inspired back‐off selection (PBS) technique. The proposed technique works on the hypothesis that synchronous collisions of beacons transmitted by a subject vehicle can be reduced if all its neighbors select different back‐off slots (ie, not the back‐off slot selected by the subject vehicle). The discrete‐event simulations demonstrate that PBS can increase the overall message reception from a subject vehicle, in comparison with the uniform random probability back‐off selection in IEEE 802.11p.     

A Modular Vaccine Platform Combining Self‐Assembled Peptide Cages and Immunogenic Peptides

Subunit vaccines use delivery platforms to present minimal antigenic components for immunization. The benefits of such systems include multivalency, self‐adjuvanting properties, and more specific immune responses. Previously, the design, synthesis, and characterization of self‐assembling peptide cages (SAGEs) have been reported. In these, de novo peptides are combined to make hubs that assemble into nanoparticles when mixed in aqueous solution. Here it is shown that SAGEs are nontoxic particles with potential as accessible synthetic peptide scaffolds for the delivery of immunogenic components. To this end, SAGEs functionalized with the model antigenic peptides tetanus toxoid_632‐651 and ovalbumin_323‐339 drive antigen‐specific responses both in vitro and in vivo, eliciting both CD4⁺ T cell and B cell responses. Additionally, SAGEs functionalized with the antigenic peptide hemagglutinin_518‐526 from the influenza virus are also able to drive a CD8⁺ T cell response in vivo. This work demonstrates the potential of SAGEs to act as a modular scaffold for antigen delivery, capable of inducing and boosting specific and tailored immune responses.     

Production of sustainable fibre-reinforced concrete incorporating waste chopped metallic film fibres and palm oil fuel ash

The consumption of waste materials is one of the essential concerns of waste management strategies in many parts of the world. With the advances in concrete technology, the utilisation of waste materials in the sustainable construction has developed increasingly widespread because of technological, economic and ecological advantages. This paper presents the workability and mechanical properties of concrete incorporating waste chopped metallic film (WCMF) fibres and palm oil fuel ash (POFA). Waste plastic results in waste discarding disaster and consequently causes significant harms to the environment. WCMF fibres were prepared by recycling metallic–plastic films used for food packaging. Six concrete mixes containing 0–1.25% WCMF fibres with a length of 20 mm were made of ordinary Portland cement (OPC). Further, six concrete mixes with the same fibre content were made, where 20% POFA substituted OPC. The combination of WCMF fibres and POFA decreased the workability of concrete mixes. The inclusion of WCMF fibres to OPC and POFA concrete mixes decreased the compressive strength. However, at the curing period of 91 days, the POFA-based mixes obtained higher compressive strength values than those of OPC-based mixtures. The positive interaction between WCMF fibres and POFA consequently enhanced the flexural and tensile strengths, impact resistance, thereby increasing energy absorption capacity and ductility of concrete composites. It revealed that WCMF fibres acted as a bridge arrester and improved the load-transfer capacity of the concrete specimens. The study showed that the utilisation of WCMF fibres in the production of sustainable concrete is a beneficial, affordable and feasible solution.     

Preventing overpacking of compressed moderately dense granular gases

Granular Matter - Predicting the force exerted on an object as it penetrates a granular medium is of interest in engineering, locomotive, and geotechnical applications. Current models of granular...     

Recognition of prosody following unilateral brain lesion: influence of functional and structural attributes of prosodic contours

The perception of prosodic distinctions by adults with unilateral right- (RHD) and left-hemisphere (LHD) damage and subjects without brain injury was assessed through six tasks that varied both functional (i.e. linguistic/emotional) and structural (i.e. acoustic) attributes of a common set of base stimuli. Three tasks explored the subjects' ability to perceive local prosodic markers associated with emphatic stress (Focus Perception condition) and three tasks examined the comprehension of emotional-prosodic meanings by the same listeners (Emotion Perception condition). Within each condition, an initial task measured the subjects' ability to recognize each "type" of prosody when all potential acoustic features (but no semantic features) signalled the target response (Baseline). Two additional tasks investigated the extent to which each group's performance on the Baseline task was influenced by duration (D-Neutral) or fundamental frequency (F-Neutral) parameters of the stimuli within each condition. Results revealed that both RHD and LHD patients were impaired, relative to healthy control subjects, in interpreting the emotional meaning of prosodic contours, but that only LHD patients displayed subnormal capacity to perceive linguistic (emphatic) specifications via prosodic cues. The performance of the RHD and LHD patients was also selectively disturbed when certain acoustic properties of the stimuli were manipulated, suggesting that both functional and structural attributes of prosodic patterns may be determinants of prosody lateralization.     

The "8-kD" cytoplasmic dynein light chain is required for nuclear migration and for dynein heavy chain localization in Aspergillus nidulans

The heavy chain of cytoplasmic dynein is required for nuclear migration in Aspergillus nidulans and other fungi. Here we report on a new gene required for nuclear migration, nudG, which encodes a homologue of the "8-kD" cytoplasmic dynein light chain (CDLC). We demonstrate that the temperature sensitive nudG8 mutation inhibits nuclear migration and growth at restrictive temperature. This mutation also inhibits asexual and sexual sporulation, decreases the intracellular concentration of the nudG CDLC protein and causes the cytoplasmic dynein heavy chain to be absent from the mycelial tip, where it is normally located in wild-type mycelia. Coimmunoprecipitation experiments with antibodies against the cytoplasmic dynein heavy chain (CDHC) and the nudG CDLC demonstrated that some fraction of the cytoplasmic dynein light chain is in a protein complex with the CDHC. Sucrose gradient sedimentation analysis, however, showed that not all of the NUDG protein is complexed with the heavy chain. A double mutant carrying a cytoplasmic dynein heavy chain deletion plus a temperature-sensitive nudG mutation grew no more slowly at restrictive temperature than a strain with only the CDHC deletion. This result demonstrates that the effect of the nudG mutation on nuclear migration and growth is mediated through an interaction with the CDHC rather than with some other molecule (e.g., myosin-V) with which the 8-kD CDLC might theoretically interact.     

Cardiovascular risk factors and educational attainment in Southern Spain: a study of a random sample of 3091 adults

Muscle mass decreases with age, leading to "sarcopenia," or low relative muscle mass, in elderly people. Sarcopenia is believed to be associated with metabolic, physiologic, and functional impairments and disability. Methods of estimating the prevalence of sarcopenia and its associated risks in elderly populations are lacking. Data from a population-based survey of 883 elderly Hispanic and non-Hispanic white men and women living in New Mexico (the New Mexico Elder Health Survey, 1993-1995) were analyzed to develop a method for estimating the prevalence of sarcopenia. An anthropometric equation for predicting appendicular skeletal muscle mass was developed from a random subsample (n = 199) of participants and was extended to the total sample. Sarcopenia was defined as appendicular skeletal muscle mass (kg)/height2 (m2) being less than two standard deviations below the mean of a young reference group. Prevalences increased from 13-24% in persons under 70 years of age to >50% in persons over 80 years of age, and were slightly greater in Hispanics than in non-Hispanic whites. Sarcopenia was significantly associated with self-reported physical disability in both men and women, independent of ethnicity, age, morbidity, obesity, income, and health behaviors. This study provides some of the first estimates of the extent of the public health problem posed by sarcopenia.