Age-associated changes in protein oxidation and proteasome activities in rat brain: modulation by antioxidants

The free radical theory of ageing postulates that age-associated neurodegeneration is caused by an imbalance between pro-oxidants and antioxidants resulting in oxidative stress. The current study showed regional variation in brain susceptibility to age-associated oxidative stress as shown by increased lipofuscin deposition and protein carbonyl levels in male rats of age 15-16 months compared to control ones (3-5 months). The hippocampus is the area most vulnerable to change compared to the cortex and cerebellum. However, proteasomal enzyme activity was not affected by age in any of the brain regions studied. Treatment with melatonin or coenzyme Q10 for 4 weeks reduced the lipofuscin content of the hippocampus and carbonyl level. However, both melatonin and coenzyme Q10 treatments inhibited beta-glutamyl peptide hydrolase activity. This suggests that these molecules can alter proteasome function independently of their antioxidant actions. (c) 2005 Elsevier Inc. All rights reserved.

Dynamic behavior of Arabidopsis eif4a-iii, putative core protein of exon junction complex: fast relocation to nucleolus and splicing speckles under hypoxia

Here, we identify the Arabidopsis thaliana ortholog of the mammalian DEAD box helicase, eIF4A-III, the putative anchor protein of exon junction complex (EJC) on mRNA. Arabidopsis eIF4A-III interacts with an ortholog of the core EJC component, ALY/Ref, and colocalizes with other EJC components, such as Mago, Y14, and RNPS1, suggesting a similar function in EJC assembly to animal eIF4A-III. A green fluorescent protein (GFP)-eIF4A-III fusion protein showed localization to several subnuclear domains: to the nucleoplasm during normal growth and to the nucleolus and splicing speckles in response to hypoxia. Treatment with the respiratory inhibitor sodium azide produced an identical response to the hypoxia stress. Treatment with the proteasome inhibitor MG132 led to accumulation of GFP-eIF4A-III mainly in the nucleolus, suggesting that transition of eIF4A-III between subnuclear domains and/or accumulation in nuclear speckles is controlled by proteolysis-labile factors. As revealed by fluorescence recovery after photobleaching analysis, the nucleoplasmic fraction was highly mobile, while the speckles were the least mobile fractions, and the nucleolar fraction had an intermediate mobility. Sequestration of eIF4A-III into nuclear pools with different mobility is likely to reflect the transcriptional and mRNA processing state of the cell.

Flow structure and variability in the subtropical Indian Ocean: Instability of the South Indian Ocean Countercurrent

The origin of the eddy variability around the 25°S band in the Indian Ocean is investigated. We have found that the surface circulation east of Madagascar shows an anticyclonic subgyre bounded to the south by eastward flow from southwest Madagascar, and to the north by the westward flowing South Equatorial Current (SEC) between 15° and 20°S. The shallow, eastward flowing South Indian Ocean Countercurrent (SICC) extends above the deep reaching, westward flowing SEC to 95°E around the latitude of the high variability band. Applying a two-layer model reveals that regions of large vertical shear along the SICC-SEC system are baroclinically unstable. Estimates of the frequencies (3.5–6 times/year) and wavelengths (290–470 km) of the unstable modes are close to observations of the mesoscale variability derived from altimetry data. It is likely then that Rossby wave variability locally generated in the subtropical South Indian Ocean by baroclinic instability is the origin of the eddy variability around 25°S as seen, for example, in satellite altimetry.

Effect of prolonged intravenous glucose and essential amino acid infusion on nitrogen balance, muscle protein degradation and ubiquitin-conjugating enzyme gene expression in calves

Background: Intravenous infusions of glucose and amino acids increase both nitrogen balance and muscle accretion. We hypothesised that co-infusion of glucose ( to stimulate insulin) and essential amino acids (EAA) would act additively to improve nitrogen balance by decreasing muscle protein degradation in association with alterations in muscle expression of components of the ubiquitin-proteasome proteolytic pathway. Methods: We examined the effect of a 5 day intravenous infusions of saline, glucose, EAA and glucose + EAA, on urinary nitrogen excretion and muscle protein degradation. We carried out the study in 6 restrained calves since ruminants offer the advantage that muscle protein degradation can be assessed by excretion of 3 methyl-histidine and multiple muscle biopsies can be taken from the same animal. On the final day of infusion blood samples were taken for hormone and metabolite measurement and muscle biopsies for expression of ubiquitin, the 14-kDa E2 ubiquitin conjugating enzyme, and proteasome sub-units C2 and C8. Results: On day 5 of glucose infusion, plasma glucose, insulin and IGF-1 concentrations were increased while urea nitrogen excretion and myofibrillar protein degradation was decreased. Co-infusion of glucose + EAA prevented the loss of urinary nitrogen observed with EAA infusions alone and enhanced the increase in plasma IGF-1 concentration but there was no synergistic effect of glucose + EAA on the decrease in myofibrillar protein degradation. Muscle mRNA expression of the ubiquitin conjugating enzyme, 14-kDa E2 and proteasome sub-unit C2 were significantly decreased, after glucose but not amino acid infusions, and there was no further response to the combined infusions of glucose + EAA. Conclusion: Prolonged glucose infusion decreases myofibrillar protein degradation, prevents the excretion of infused EAA, and acts additively with EAA to increase plasma IGF-1 and improve net nitrogen balance. There was no evidence of synergistic effects between glucose + EAA infusion on muscle protein degradation or expression of components of the ubiquitin-proteasome proteolytic pathway.

PDGF regulates the actin cytoskeleton through hnRNP-K-mediated activation of the ubiquitin E-3-ligase MIR

PDGF is a potent chemotactic mitogen and a strong inductor of fibroblast motility. In Swiss 3T3 fibroblasts, exposure to PDGF but not EGF or IGF-1 causes a rapid loss of actin stress fibers (SFs) and focal adhesions (FAs), which is followed by the development of retractile dendritic protrusions and induction of motility. The PDGF-specific actin reorganization was blocked by inhibition of Src-kinase and the 26S proteasome. PDGF induced Src-dependent association between the multifunctional transcription/translation regulator hnRNP-K and the mRNA-encoding myosin regulatory light-chain (MRLC)-interacting protein (MIR), a E3-ubiquitin ligase that is MRLC specific. This in turn rapidly increased MIR expression, and led to ubiquitination and proteasome-mediated degradation of MRLC. Downregulation of MIR by RNA muting prevented the reorganization of actin structures and severely reduced the migratory and wound-healing potential of PDGF-treated cells. The results show that activation of MIR and the resulting removal of diphosphorylated MRLC are essential for PDGF to instigate and maintain control over the actin-myosin-based contractile system in Swiss 3T3 fibroblasts. The PDGF induced protein destabilization through the regulation of hnRNP-K controlled ubiquitin-ligase translation identifies a novel pathway by which external stimuli can regulate phenotypic development through rapid, organelle-specific changes in the activity and stability of cytoskeletal regulators.

Occupational choice, socio-economic status and educational attainment: a study of the occupational choices and destinations of young people in the British household panel survey

The article considers young people's occupational choices at the age of 15 in relation to their educational attainment, the occupations of their parents and their actual occupations when they are in their early 20s. It uses data from the British Household Panel Survey over periods of between five and ten years. The young people in the survey are occupationally ambitious: many more aspire to professional, managerial and technical jobs than the likely availability of these occupations. In general ambitions and educational attainment and intentions are well aligned but there are also many instances of misalignment; either people wanting jobs which their educational attainments and intentions will not prepare them for, or people with less ambitious aspirations than their educational performance would justify. Children from more occupationally advantaged families are more ambitious, achieve better educationally and have better occupational outcomes than other children. However, where young people are both ambitious and educationally successful the occupational outcomes are as good for those from disadvantaged as advantaged families. In contrast, where young people are neither ambitious nor educationally successful, the outcomes for those from disadvantaged homes are very much poorer than for other young people. The article suggests that while choice is real it is also heavily constrained for many people. A possible educational implication of the study is that career interventions could be directed at under-ambitious but academically capable young people from disadvantaged backgrounds.

Upper-ocean heat budget and ocean eddy transport in the south-east Pacific in a high-resolution coupled model

We present an analysis of the oceanic heat advection and its variability in the upper 500 m in the southeastern tropical Pacific (100W–75W, 25S–10S) as simulated by the global coupled model HiGEM, which has one of the highest resolutions currently used in long-term integrations. The simulated climatology represents a temperature advection field arising from transient small-scale (<450 km) features, with structures and transport that appear consistent with estimates based on available observational data for the mooring at 20S, 85W. The transient structures are very persistent (>4 months), and in specific locations they generate an important contribution to the local upper-ocean heat budget, characterised by scales of a few hundred kilometres, and periods of over a year. The contribution from such structures to the local, long-term oceanic heat budget however can be of either sign, or vanishing, depending on the location; and, although there appears some organisation in preferential areas of activity, the average over the entire region is small. While several different mechanisms may be responsible for the temperature advection by transients, we find that a significant, and possibly dominant, component is associated with vortices embedded in the large-scale, climatological salinity gradient associated with the fresh intrusion of mid-latitude intermediate water which penetrates north-westward beneath the tropical thermocline

20th century intraseasonal Asian monsoon dynamics viewed from Isomap

The Asian summer monsoon is a high dimensional and highly nonlinear phenomenon involving considerable moisture transport towards land from the ocean, and is critical for the whole region. We have used daily ECMWF reanalysis (ERA-40) sea-level pressure (SLP) anomalies to the seasonal cycle, over the region 50-145°E, 20°S-35°N to study the nonlinearity of the Asian monsoon using Isomap. We have focused on the two-dimensional embedding of the SLP anomalies for ease of interpretation. Unlike the unimodality obtained from tests performed in empirical orthogonal function space, the probability density function, within the two-dimensional Isomap space, turns out to be bimodal. But a clustering procedure applied to the SLP data reveals support for three clusters, which are identified using a three-component bivariate Gaussian mixture model. The modes are found to appear similar to active and break phases of the monsoon over South Asia in addition to a third phase, which shows active conditions over the Western North Pacific. Using the low-level wind field anomalies the active phase over South Asia is found to be characterised by a strengthening and an eastward extension of the Somali jet whereas during the break phase the Somali jet is weakened near southern India, while the monsoon trough in northern India also weakens. Interpretation is aided using the APHRODITE gridded land precipitation product for monsoon Asia. The effect of large-scale seasonal mean monsoon and lower boundary forcing, in the form of ENSO, is also investigated and discussed. The outcome here is that ENSO is shown to perturb the intraseasonal regimes, in agreement with conceptual ideas.

Exploring invention capability

Research on invention has focused on business invention and little work has been conducted on the process and capability required for the individual inventor or the capabilities required for an advice to be considered an invention. This paper synthesises the results of an empirical survey of ten inventor case studies with current research on invention and recent capability affordance research to develop an integrated capability process model of human capabilities for invention and specific capabilities of an invented device. We identify eight necessary human effectivities required for individual invention capability and six functional key activities using these effectivities, to deliver the functional capability of invention. We also identified key differences between invention and general problem solving processes. Results suggest that inventive step capability relies on a unique application of principles that relate to a new combination of affordance chain with a new mechanism and or space time (affordance) path representing the novel way the device works, in conjunction with defined critical affordance operating factors that are the subject of the patent claims.

Mycolactone-dependent depletion of endothelial cell thrombomodulin is strongly associated with fibrin deposition in buruli ulcer lesions

A well-known histopathological feature of diseased skin in Buruli ulcer (BU) is coagulative necrosis caused by the Mycobacterium ulcerans macrolide exotoxin mycolactone. Since the underlying mechanism is not known, we have investigated the effect of mycolactone on endothelial cells, focussing on the expression of surface anticoagulant molecules involved in the protein C anticoagulant pathway. Congenital deficiencies in this natural anticoagulant pathway are known to induce thrombotic complications such as purpura fulimans and spontaneous necrosis. Mycolactone profoundly decreased thrombomodulin (TM) expression on the surface of human dermal microvascular endothelial cells (HDMVEC) at doses as low as 2ng/ml and as early as 8hrs after exposure. TM activates protein C by altering thrombin’s substrate specificity, and exposure of HDMVEC to mycolactone for 24 hours resulted in an almost complete loss of the cells’ ability to produce activated protein C. Loss of TM was shown to be due to a previously described mechanism involving mycolactone-dependent blockade of Sec61 translocation that results in proteasome-dependent degradation of newly synthesised ER-transiting proteins. Indeed, depletion from cells determined by live-cell imaging of cells stably expressing a recombinant TM-GFP fusion protein occurred at the known turnover rate. In order to determine the relevance of these findings to BU disease, immunohistochemistry of punch biopsies from 40 BU lesions (31 ulcers, nine plaques) was performed. TM abundance was profoundly reduced in the subcutis of 78% of biopsies. Furthermore, it was confirmed that fibrin deposition is a common feature of BU lesions, particularly in the necrotic areas. These findings indicate that there is decreased ability to control thrombin generation in BU skin. Mycolactone’s effects on normal endothelial cell function, including its ability to activate the protein C anticoagulant pathway are strongly associated with this. Fibrin-driven tissue ischemia could contribute to the development of the tissue necrosis seen in BU lesions.

Predicting fatty acid profiles in blood based on food intake and the FADS1 rs174546 SNP

SCOPE: A high intake of n-3 PUFA provides health benefits via changes in the n-6/n-3 ratio in blood. In addition to such dietary PUFAs, variants in the fatty acid desaturase 1 (FADS1) gene are also associated with altered PUFA profiles. METHODS AND RESULTS: We used mathematical modelling to predict levels of PUFA in whole blood, based on MHT and bolasso selected food items, anthropometric and lifestyle factors, and the rs174546 genotypes in FADS1 from 1,607 participants (Food4Me Study). The models were developed using data from the first reported time point (training set) and their predictive power was evaluated using data from the last reported time point (test set). Amongst other food items, fish, pizza, chicken and cereals were identified as being associated with the PUFA profiles. Using these food items and the rs174546 genotypes as predictors, models explained 26% to 43% of the variability in PUFA concentrations in the training set and 22% to 33% in the test set. CONCLUSIONS: Selecting food items using MHT is a valuable contribution to determine predictors, as our models' predictive power is higher compared to analogue studies. As unique feature, we additionally confirmed our models' power based on a test set.

An assessment of air-sea heat fluxes from ocean and coupled reanalyses

Sixteen monthly air–sea heat flux products from global ocean/coupled reanalyses are compared over 1993–2009 as part of the Ocean Reanalysis Intercomparison Project (ORA-IP). Objectives include assessing the global heat closure, the consistency of temporal variability, comparison with other flux products, and documenting errors against in situ flux measurements at a number of OceanSITES moorings. The ensemble of 16 ORA-IP flux estimates has a global positive bias over 1993–2009 of 4.2 ± 1.1 W m−2. Residual heat gain (i.e., surface flux + assimilation increments) is reduced to a small positive imbalance (typically, +1–2 W m−2). This compensation between surface fluxes and assimilation increments is concentrated in the upper 100 m. Implied steady meridional heat transports also improve by including assimilation sources, except near the equator. The ensemble spread in surface heat fluxes is dominated by turbulent fluxes (>40 W m−2 over the western boundary currents). The mean seasonal cycle is highly consistent, with variability between products mostly <10 W m−2. The interannual variability has consistent signal-to-noise ratio (~2) throughout the equatorial Pacific, reflecting ENSO variability. Comparisons at tropical buoy sites (10°S–15°N) over 2007–2009 showed too little ocean heat gain (i.e., flux into the ocean) in ORA-IP (up to 1/3 smaller than buoy measurements) primarily due to latent heat flux errors in ORA-IP. Comparisons with the Stratus buoy (20°S, 85°W) over a longer period, 2001–2009, also show the ORA-IP ensemble has 16 W m−2 smaller net heat gain, nearly all of which is due to too much latent cooling caused by differences in surface winds imposed in ORA-IP.