Increased expression of phosphorylated forms of RNA-dependent protein kinase and eukaryotic initiation factor 2 alpha may signal skeletal muscle atrophy in weight-losing cancer patients

Previous studies suggest that the activation (autophosphorylation) of dsRNA-dependent protein kinase (PKR) can stimulate protein degradation, and depress protein synthesis in skeletal muscle through phosphorylation of the translation initiation factor 2 (eIF2) on the alpha-subunit. To understand whether these mediators are important in muscle wasting in cancer patients, levels of the phospho forms of PKR and eIF2alpha have been determined in rectus abdominus muscle of weight losing patients with oesophago-gastric cancer, in comparison with healthy controls. Levels of both phospho PKR and phospho eIF2alpha were significantly enhanced in muscle of cancer patients with weight loss irrespective of the amount and there was a linear relationship between phosphorylation of PKR and phosphorylation of eIF2alpha (correlation coefficient 0.76, P=0.005). This suggests that phosphorylation of PKR led to phosphorylation of eIF2alpha. Myosin levels decreased as the weight loss increased, and there was a linear relationship between myosin expression and the extent of phosphorylation of eIF2alpha (correlation coefficient 0.77, P=0.004). These results suggest that phosphorylation of PKR may be an important initiator of muscle wasting in cancer patients.

Initiation and progression of ossification of the posterior longitudinal ligament of the cervical spine in the hereditary spinal hyperostotic mouse (twy/twy)

Ossification of the posterior longitudinal ligament (OPLL) is a significantly critical pathology that can eventually cause serious myelopathy. Ossification commences in the vertebral posterior longitudinal ligaments, and intensifies and spreads with the progression of the disease, resulting in osseous projections and compression of the spinal cord. However, the paucity of histological studies the underlying mechanisms of calcification and ossification processes remain obscure. The pathological process could be simulated in the ossifying process of the ligament in mutant spinal hyperostotic mouse (twy/twy). The aim of this study is to observe that enlargement of the nucleus pulposus followed by herniation, disruption and regenerative proliferation of annulus fibrosus cartilaginous tissues participated in the initiation of ossification of the posterior longitudinal ligament of twy/twy mice.

Rapid depletion of mutant eukaryotic initiation factor 5A at restrictive temperature reveals connections to actin cytoskeleton and cell cycle progression

Eukaryotic initiation factor 5A (eIF5A) is the only protein in nature that contains hypusine, an unusual amino acid derived from the modification of lysine by spermidine. Two genes, TIF51A and TIF51B, encode eIF5A in the yeast Saccharomyces cerevisiae. In an effort to understand the structure-function relationship of eIF5A, we have generated yeast mutants by introducing plasmid-borne tif51A into a double null strain where both TIF51A and TIF51B have been disrupted. One of the mutants, tsL102A strain (tif51A L102A tif51aDelta tif51bDelta) exhibits a strong temperature-sensitive growth phenotype. At the restrictive temperature, tsL102A strain also exhibits a cell shape change, a lack of volume change in response to temperature increase and becomes more sensitive to ethanol, a hallmark of defects in the PKC/WSC cell wall integrity pathway. In addition, a striking change in actin dynamics and a complete cell cycle arrest at G1 phase occur in tsL102A cells at restrictive temperature. The temperature-sensitivity of tsL102A strain is due to a rapid loss of mutant eIF5A with the half-life reduced from 6 h at permissive temperature to 20 min at restrictive temperature. Phenylmethyl sulfonylfluoride (PMSF), an irreversible inhibitor of serine protease, inhibited the degradation of mutant eIF5A and suppressed the temperature-sensitive growth arrest. Sorbitol, an osmotic stabilizer that complement defects in PKC/WSC pathways, stabilizes the mutant eIF5A and suppresses all the observed temperature-sensitive phenotypes.

Novel characteristics of the biological properties of the yeast Saccharomyces cerevisiae eukaryotic initiation factor 2A

Eukaryotic initiation factor 2A (eIF2A) has been shown to direct binding of the initiator methionyl-tRNA (Met-tRNA(i)) to 40 S ribosomal subunits in a codon-dependent manner, in contrast to eIF2, which requires GTP but not the AUG codon to bind initiator tRNA to 40 S subunits. We show here that yeast eIF2A genetically interacts with initiation factor eIF4E, suggesting that both proteins function in the same pathway. The double eIF2A/eIF4E-ts mutant strain displays a severe slow growth phenotype, which correlated with the accumulation of 85% of the double mutant cells arrested at the G(2)/M border. These cells also exhibited a disorganized actin cytoskeleton and elevated actin levels, suggesting that eIF2A might be involved in controlling the expression of genes involved in morphogenic processes. Further insights into eIF2A function were gained from the studies of eIF2A distribution in ribosomal fractions obtained from either an eIF5BDelta (fun12Delta) strain or a eIF3b-ts (prt1-1) strain. It was found that the binding of eIF2A to 40 and 80 S ribosomes was not impaired in either strain. We also found that eIF2A functions as a suppressor of Ure2p internal ribosome entry site-mediated translation in yeast cells. The regulation of expression from the URE2 internal ribosome entry site appears to be through the levels of eIF2A protein, which has been found to be inherently unstable with a half-life of approximately 17 min. It was hypothesized that this instability allows for translational control through the level of eIF2A protein in yeast cells.

Fatigue of commercial aluminium alloys

Fatigue crack propagation has been observed for a number of commercial aluminium alloys. Comparable data was obtained for a variety of crack and specimen geometries over a range of crack lengths for a given alloy. Where crack propagation only was of interest the initiation event has been excluded by pre-cracking the specimen using a fin of material adjacent to the crack face. By this method a controlled defect size is introduced in to the specimen. By modification of the D.C. potential drop method it has been shown possible to measure the growth of cracking from 0.12mm by this method. Crack growth from defects greater than 0.6mm have been shown to give conventional crack propagation deduced by principle of similitude. Fatigue fracture surface analysis has been conducted for cracking from both free surfaces and from blunt notches. A `quasi cleavage' feature has been identified and is shown to be prominent when the fatigue stress intensity range is below 10 MNm-3/2.

"Crack down on the celebrity junkies": does media coverage of celebrity drug use pose a risk to young people?

This study analysed news media content to examine the role played by celebrity drug use in young people's perceptions of drug use. We know that young people have access to discourses of drug use through music and other media which may emphasise short term gains (of pleasure or sexual success) over longer term health and social problems. This study goes beyond a simple modelling approach by using Media Framing Analysis (MFA) to take an in-depth look at the messages themselves and how they are 'framed'. New stories about Amy Winehouse's drug use were used and we conducted focus groups with young people asking them questions about drugs, celebrity and the media. Frames identified include: 'troubled genius', 'losing patience' and 'glamorization or gritty realism'. Initially, the press championed Winehouse's musical talent but soon began to tire of her recklessness; the participants tended to be unimpressed with Winehouse's drug use, characterising her as a promising artist who had 'gone off the rails'. Young people were far more critical of Winehouse than might be expected, demonstrating that concerns about the influence of celebrity drug use and its impact on future health risk behaviour among young people may have been over-simplified and exaggerated. This study illustrates the need to understand young people and their frames of reference within popular culture when designing drug awareness information relevant to them. Furthermore, it indicates that critical media skills analysis may contribute to health risk education programmes related to drug use.

Eukaryotic translation initiation factor 3, subunit a, regulates the extracellular signal-regulated kinase pathway

The extracellular signal-regulated kinase (ERK) pathway participates in the control of numerous cellular processes, including cell proliferation. Since its activation kinetics are critical for to its biological effects, they are tightly regulated. We report that the protein translation factor, eukaryotic translation initiation factor 3, subunit a (eIF3a), binds to SHC and Raf-1, two components of the ERK pathway. The interaction of eIF3a with Raf-1 is increased by ß-arrestin2 expression and transiently decreased by epidermal growth factor (EGF) stimulation in a concentration-dependent manner. The EGF-induced decrease in Raf-1-eIF3a association kinetically correlates with the time course of ERK activation. eIF3a interferes with Raf-1 activation and eIF3a downregulation by small interfering RNA enhances ERK activation, early gene expression, DNA synthesis, expression of neuronal differentiation markers in PC12 cells, and Ras-induced focus formation in NIH 3T3 cells. Thus, eIF3a is a negative modulator of ERK pathway activation and its biological effects.

Crack-growth of medical-grade silicone using pure shear tests

Silicone elastomers are commonly used in the manufacture of single-piece joint replacement implants for the finger joints. However, the survivorship of these implants can be poor, with failure typically occurring from fracture of the stems. The aim of this paper was to investigate the crack growth of medical-grade silicone using pure shear tests. Two medical-grade silicones (C6-180 and Med82-5010-80) were tested. Each sample had a 20 mm crack introduced and was subjected to a sinusoidally varying tensile strain, with a minimum of 0 per cent and a maximum in the range 10 to 77 per cent. Testing was undertaken at a frequency of 10 Hz. At various times during testing, the testing machine was stopped, the number of cycles completed was noted, and the crack length measured. Graphs of crack length against number of cycles were plotted, as well as the crack growth rate against tearing energy. The results show that Med82-5010-80 is more crack resistant than C6-180. Graphs of crack growth rate against tearing energy can be used to predict the failure of these medical-grade elastomers.

H-induced platelet and crack formation in hydrogenated epitaxial Si/Si0.98 B0.02 /Si structures

An approach to transfer a high-quality Si layer for the fabrication of silicon-on-insulator wafers has been proposed based on the investigation of platelet and crack formation in hydrogenated epitaxialSi/Si0.98B0.02/Si structures grown by molecular-beam epitaxy. H-related defect formation during hydrogenation was found to be very sensitive to the thickness of the buried Si0.98B0.02 layer. For hydrogenated Si containing a 130nm thick Si0.98B0.02 layer, no platelets or cracking were observed in the B-doped region. Upon reducing the thickness of the buried Si0.98B0.02 layer to 3nm, localized continuous cracking was observed along the interface between the Si and the B-doped layers. In the latter case, the strains at the interface are believed to facilitate the (100)-oriented platelet formation and (100)-oriented crack propagation.

Fatigue crack propagation in some PM steels

Mechanisms of fatigue crack growth have been studied for a range of PM steels at relative densities of 0.90 and 1.0, for which strength, fracture toughness, and microstructural information was also available. It is shown that the Paris exponents for steady state crack growth are between 8 and 18 when ρr is approximately 0.9 but when ρr is approximately 1.0 the exponents are between 2.6 and 4.0, i.e in the range typical of wrought steels (2-4). At both densities, threshold stress intensities are between 5.5 and 10.8 MPa m1/2 when R = 0.1. Combinations of these thresholds and yield strengths are comparable with those for wrought steels. When R = 0.8, reductions in threshold to between 2.7 and 5 MPa m1/2 are attributed to crack closure effects. At ρr = 0.90, Fe-0.5C fails by progressive rupture of sinter necks. Astaloy A, with 0.2%C and 0.6%C, and Distaloy AB-0.6C have smaller plastic zone sizes and the cracks follow more difficult paths through particles as well as necks. When ρr is approximately 1.0, fracture is partially by true fatigue modes and partly by cleavage, the bursts of cleavage being more noticeable when Kmax is high.

The effect of frequency and microstructure on corrosion fatigue crack propagation in high strength aluminium alloys

Fatigue crack growth in high strength aluminium alloy 7150 commercial plate material has been studied in both laboratory air and acidified aqueous salt solution. The aggressive aqueous environment enhanced fatigue crack growth rates by up to an order in magnitude compared to laboratory air. The enhancement in fatigue crack growth rate was accompanied by evidence of embrittlement in the crack path, involving both brittle intergranular and transgranular failure modes. Both the enhancement of fatigue crack growth rates and the extent of intergranular growth modes are dependent on cyclic frequency which, along with the absence of a similar frequency effect in a spray-formed version of the material with a significantly different grain structure, supports a mechanism of grain boundary hydrogen diffusion for intergranular corrosion fatigue crack growth. The convergence of corrosion fatigue crack growth rates at high ΔK in both spray-formed and conventional plate materials coincides with the operation of identical transgranular corrosion fatigue modes dependent on strain-controlled hydrogen diffusion ahead of the crack tip. © 1997 Acta Metallurgica Inc.

Isothermal fatigue of an aluminide-coated single-crystal superalloy:Part II. Effects of brittle precracking

The effect of brittle coating precracking on the fatigue behavior of a high-activity aluminide-coated single-crystal nickel-base superalloy has been studied using hollow cylindrical specimens at test temperatures of 600 °C, 800 °C, and 1000 °C. Three types of precrack were studied: narrow precracks formed at room temperature, wide precracks formed at room temperature, and narrow precracks formed at elevated temperature. The effect of precracking on fatigue life at 600 °C was found to depend strongly on the type of precrack. No failure was observed for specimens with narrow room-temperature precracks because of crack arrest via an oxidation-induced crack closure mechanism, while the behavior of wide precracks and precracks formed at elevated temperature mirrored the non-precracked behavior. Crack retardation also occurred for narrow room-temperature precracks tested at 800 °C - in this case, fatigue cracks leading to failure initiated in a layer of recrystallized grains on the inside surface of the specimen. A significant reduction in fatigue life at 800 °C relative to non-precracked specimens was observed for wide precracks and elevated temperature precracks. The presence of precracks bypassed the initiation and growth of coating fatigue cracks necessary for failure in non-precracked material. No effect of precracking was observed at 1000 °C.