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.

The strength of column-to-foundation joints in reinforced concrete

This thesis is concerned with the experimental and theoretical investigation into the compression bond of column longitudinal reinforcement in the transference of axial load from a reinforced concrete column to a base. Experimental work includes twelve tests with square twisted bars and twenty four tests with ribbed bars. The effects of bar size, anchorage length in the base, plan area of the base, provision of bae tensile reinforcement, links around the column bars in the base, plan area of column and concrete compressive strength were investigated in the tests. The tests indicated that the strength of the compression anchorage of deformed reinforcing steel in the concrete was primarily dependent on the concrete strength and the resistance to bursting, which may be available within the anchorage . It was shown in the tests without concreted columns that due to a large containment over the bars in the foundation, failure occurred due to the breakdown of bond followed by the slip of the column bars along the anchorage length. The experimental work showed that the bar size , the stress in the bar, the anchorage length, provision of the transverse steel and the concrete compressive strength significantly affect the bond stress at failure. The ultimate bond stress decreases as the anchorage length is increased, while the ultimate bond stress increases with increasing each of the remainder parameters. Tests with concreted columns also indicated that a section of the column contributed to the bond length in the foundation by acting as an extra anchorage length. The theoretical work is based on the Mindlin equation( 3), an analytical method used in conjunction with finite difference calculus. The theory is used to plot the distribution of bond stress in the elastic and the elastic-plastic stage of behaviour. The theory is also used to plot the load-vertical displacement relationship of the column bars in the anchorage length, and also to determine the theoretical failure load of foundation. The theoretical solutions are in good agreement with the experimental results and the distribution of bond stress is shown to be significantly influenced by the bar stiffness factor K. A comparison of the experimental results with the current codes shows that the bond stresses currently used are low and in particular, CPIlO(56) specifies very conservative design bond stresses .

A Tale of Two Fishes: Using Recreational Angler Records to Examine the Link Between Fish Catches and Floodplain Connections in a Subtropical Coastal River

In the tropical and subtropical wet and dry regions, maintaining natural hydrologic connections between coastal rivers and adjacent ephemeral wetlands is critical to conserving and sustaining high levels of fisheries production within these systems. Though there is a consensus that there is a need to maintain these natural connections, little is known about what attributes of floodplain inundation regimes are most important in sustaining fisheries production. Two attributes of the flood season and thus floodplain inundation that may be particularly influential to fisheries are the amplitude of the flood season (floodplain water depth and spatial extent of inundation) and the duration of the flood season (i.e., time floodplains are inundated). In mangrove-dominated Everglades coastal rivers, seasonal inundation of upstream marsh floodplains may play an important role in provisioning recreational fisheries; however, this relationship remains unknown. Using two Everglades coastal river fisheries as a model, we tested whether the amplitude of the flood season or the duration of the flood season is more important in explaining variation in angler catch records of common snook and largemouth bass collected from 1992 to 2012. We validated angler catches with fisheries-independent electrofishing conducted in the same region from 2004 to 2012. Our results showed (1) that bass angler catches tracked electrofishing catches, while snook catches were completely mismatched. And (2) that previous year's marsh dynamics, particularly the duration of the flood season, was more influential than the flood season amplitude in explaining variation in bass catches, such that bass angler catches were negatively correlated to the period time that floodplains remained disconnected from coastal rivers in the previous year, while snook catches were not very well explained by floodplain inundation terms.

Physical oceanography, pore water chemistry and water column methane turover rates during POLARSTERN cruise ANT-XXIX/4

Recent studies have suggested that the marine contribution of methane from shallow regions and melting marine terminating glaciers may have been underestimated. Here we report on methane sources and potential sinks associated with methane seeps in Cumberland Bay, South Georgia's largest fjord system. The average organic carbon content in the upper 8 meters of the sediment is around 0.65 wt.%; this observation combined with Parasound data suggest that the methane gas accumulations probably originate from peat-bearing sediments currently located several tens of meters below the seafloor. Only one of our cores indicates upward advection; instead most of the methane is transported via diffusion. Sulfate and methane flux estimates indicate that a large fraction of methane is consumed by anaerobic oxidation of methane (AOM). Carbon cycling at the sulfate-methane transition (SMT) results in a marked fractionation of the d13C-CH4 from an estimated source value of -65 per mil to a value as low as -96 per mil just below the SMT. Methane concentrations in sediments are high, especially close to the seepage sites (~40 mM); however, concentrations in the water column are relatively low (max. 58 nM) and can be observed only close to the seafloor. Methane is trapped in the lowermost water mass, however, measured microbial oxidation rates reveal very low activity with an average turnover of 3.1 years. We therefore infer that methane must be transported out of the bay in the bottom water layer. A mean sea-air flux of only 0.005 nM/m²/s confirms that almost no methane reaches the atmosphere.

Chromatographic methods to obtain the biomolecules profile of some aromatic plants irradiated with electron beam

Irradiation is being progressively considered as a versatile and effective conservation technique [1]. Based on this premise, our research group has been investigating the effects of different irradiation conditions in several food matrices. Aromatic plants are among the food products that require suitable conservation technologies to expand their use [2]. The effects of irradiation on the four species (Aloysia citrodora, Melissa officinalis, Melittis melissophyllum and Mentha piperita) studied herein were previously evaluated. In the present study, the same species were treated with different doses of electron-beam irradiation (0, 1 and 10 kGy) and several parameters were evaluated. The individual sugars profile was determined by HPLCRI, fatty acids by GC-FID, organic acids by HPLC-PDA and tocopherols by HPLCfluorescence. In general, the evaluated parameters remained practically unchanged, regardless of plant species or the irradiation dose. Regarding the profile of sugars, the major change was a decrease in the content of disaccharides. The most notable variations in organic acids were observed in plant species with the highest content in these molecules, especially the decrease observed in the samples of M. officinalis and M. melissophyllum. Among the tocopherols, the α and β isoforms were more susceptible to radiation, while the application of 1 kGy tended to increase the levels of tocopherols in Aloysia citrodora, while 10 kGy had the same effect on M. melissophyllum. M. piperita sample showed the highest levels of tocopherols, regardless of the dose applied. Finally, with regard to the fatty acids content, the irradiated samples showed higher percentages of monounsaturated fatty acids than the control samples. In general, analyzing the results taking into account the effects described, it can be concluded that the application of irradiation with electron beam at doses 1 and 10 kGy is an effective way to retain biomolecules profile of the studied species.

Energy Loss Function of Solids Assessed by Ion Beam Energy-Loss Measurements: Practical Application to Ta2O5

We present a study where the energy loss function of Ta2O5, initially derived in the optical limit for a limited region of excitation energies from reflection electron energy loss spectroscopy (REELS) measurements, was improved and extended to the whole momentum and energy excitation region through a suitable theoretical analysis using the Mermin dielectric function and requiring the fulfillment of physically motivated restrictions, such as the f- and KK-sum rules. The material stopping cross section (SCS) and energy-loss straggling measured for 300–2000 keV proton and 200–6000 keV helium ion beams by means of Rutherford backscattering spectrometry (RBS) were compared to the same quantities calculated in the dielectric framework, showing an excellent agreement, which is used to judge the reliability of the Ta2O5 energy loss function. Based on this assessment, we have also predicted the inelastic mean free path and the SCS of energetic electrons in Ta2O5.

In situ epicatechin-loaded hydrogel implants for local drug delivery to spinal column for effective management of post-traumatic spinal injuries

Purpose: To prepare hydrogels loaded with epicatechin, a strong antioxidant, anti-inflammatory, and neuroprotective tea flavonoid, and characterise them in situ as a vehicle for prolonged and safer drug delivery in patients with post-traumatic spinal cord injury. Methods: Five in situ gel formulations were prepared using chitosan and evaluated in terms of their visual appearance, clarity, pH, viscosity, and in vitro drug release. In vivo anti-inflammatory activity was determined and compared with 2 % piroxicam gel as standard. Motor function activity in a rat model of spinal injury was examined comparatively with i.v. methylprednisolone as standard. Results: The N-methyl pyrrolidone solution (containing 1 % w/w epicatechin with 2 to 10 % w/w chitosan) of the in situ gel formulation had a uniform pH in the range of 4.01 ± 0.12 to 4.27 ± 0.02. High and uniform drug loading, ranging from 94.48 ± 1.28 to 98.08 ± 1.24 %, and good in vitro drug release (79.48 ± 2.84 to 96.48 ± 1.02 % after 7 days) were achieved. The in situ gel prepared from 1 % epicatechin and 2 % chitosan (E5) showed the greatest in vivo anti-inflammatory activity (60.58 % inhibition of paw oedema in standard carrageenan-induced hind rat paw oedema model, compared with 48.08 % for the standard). The gels showed significant therapeutic effectiveness against post-traumainduced spinal injury in rats. E5 elicited maximum motor activity (horizontal bar test) in the spinal injury rat model; the rats that received E5 treatment produced an activity score of 3.62 ± 0.02 at the end of 7 days, compared with 5.0 ± 0.20 following treatment with the standard. Conclusion: In situ epicatechin-loaded gel exhibits significant neuroprotective and anti-inflammatory effects, and therefore can potentially be used for prolonged and safe drug delivery in patients with traumatic spinal cord injury.

Beam-energy Dependence Of The Directed Flow Of Protons, Antiprotons, And Pions In Au+au Collisions.

Rapidity-odd directed flow (v1) measurements for charged pions, protons, and antiprotons near midrapidity (y=0) are reported in sNN=7.7, 11.5, 19.6, 27, 39, 62.4, and 200 GeV Au+Au collisions as recorded by the STAR detector at the Relativistic Heavy Ion Collider. At intermediate impact parameters, the proton and net-proton slope parameter dv1/dy|y=0 shows a minimum between 11.5 and 19.6 GeV. In addition, the net-proton dv1/dy|y=0 changes sign twice between 7.7 and 39 GeV. The proton and net-proton results qualitatively resemble predictions of a hydrodynamic model with a first-order phase transition from hadronic matter to deconfined matter, and differ from hadronic transport calculations.

Beam-energy Dependence Of Charge Separation Along The Magnetic Field In Au+au Collisions At Rhic.

Local parity-odd domains are theorized to form inside a quark-gluon plasma which has been produced in high-energy heavy-ion collisions. The local parity-odd domains manifest themselves as charge separation along the magnetic field axis via the chiral magnetic effect. The experimental observation of charge separation has previously been reported for heavy-ion collisions at the top RHIC energies. In this Letter, we present the results of the beam-energy dependence of the charge correlations in Au+Au collisions at midrapidity for center-of-mass energies of 7.7, 11.5, 19.6, 27, 39, and 62.4 GeV from the STAR experiment. After background subtraction, the signal gradually reduces with decreased beam energy and tends to vanish by 7.7 GeV. This implies the dominance of hadronic interactions over partonic ones at lower collision energies.