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.

Domain and nucleotide dependence of the interaction between Saccharomyces cerevisiae translation elongation factors 3 and 1A

Eukaryotic translation elongation factor 3 (eEF3) is a fungal-specific ATPase proposed to catalyze the release of deacylated-tRNA from the ribosomal E-site. In addition, it has been shown to interact with the aminoacyl-tRNA binding GTPase elongation factor 1A (eEF1A), perhaps linking the E and A sites. Domain mapping demonstrates that amino acids 775-980 contain the eEF1A binding sites. Domain III of eEF1A, which is also involved in actin-related functions, is the site of eEF3 binding. The binding of eEF3 to eEF1A is enhanced by ADP, indicating the interaction is favored post-ATP hydrolysis but is not dependent on the eEF1A-bound nucleotide. A temperature-sensitive P915L mutant in the eEF1A binding site of eEF3 has reduced ATPase activity and affinity for eEF1A. These results support the model that upon ATP hydrolysis, eEF3 interacts with eEF1A to help catalyze the delivery of aminoacyl-tRNA at the A-site of the ribosome. The dynamics of when eEF3 interacts with eEF1A may be part of the signal for transition of the post to pre-translocational ribosomal state in yeast.

In situ diagnostics and prognostics of solder fatigue in IGBT modules for electric vehicle drives

This paper proposes an in situ diagnostic and prognostic (D&P) technology to monitor the health condition of insulated gate bipolar transistors (IGBTs) used in EVs with a focus on the IGBTs' solder layer fatigue. IGBTs' thermal impedance and the junction temperature can be used as health indicators for through-life condition monitoring (CM) where the terminal characteristics are measured and the devices' internal temperature-sensitive parameters are employed as temperature sensors to estimate the junction temperature. An auxiliary power supply unit, which can be converted from the battery's 12-V dc supply, provides power to the in situ test circuits and CM data can be stored in the on-board data-logger for further offline analysis. The proposed method is experimentally validated on the developed test circuitry and also compared with finite-element thermoelectrical simulation. The test results from thermal cycling are also compared with acoustic microscope and thermal images. The developed circuitry is proved to be effective to detect solder fatigue while each IGBT in the converter can be examined sequentially during red-light stopping or services. The D&P circuitry can utilize existing on-board hardware and be embedded in the IGBT's gate drive unit.

Temperature effects on fatigue thresholds and structure sensitive crack growth in a nickel-base superalloy

Fatigue thresholds and slow crack growth rates have been measured in a powder formed nickel-base superalloy from room temperature to 600°C. Two grain sizes were investigated: 5-12 μm and 50 μm. It is shown that the threshold increases with grain size, and the difference is most pronounced at room temperature. Although crack growth rates increase with temperature in both microstructures, the threshold is only temperature dependent in the material with the larger grain size. It is also only in the latter that the room temperature threshold falls when the load ratio is increased from 0.1 to 0.5. At 600°C the higher load ratio causes a 20% reduction in the threshold irrespective of grain size. The results are discussed in terms of surface roughness and oxide-induced crack closure, the former being critically related to the type of crystallographic crack growth, which is in turn shown to be both temperature and stress intensity dependent. © 1983.

Highly sensitive temperature and strain sensors based on all-fiber 45°-TFG Lyot filter

We demonstrate highly sensitive temperature and strain sensors based on an all-fiber Lyot filter structure, which is formed by concatenating two 45°-TFGs (tilted fiber gratings) with a PM fiber cavity. The experiment results show the all-fiber 45°-TFG Lyot filter has very high sensitivity to strain and temperature. The 45°-TFG Lyot filters of two different cavity lengths (18cm and 40 cm) have been evaluated for temperature sensing by heating a section of the cavity from 10°C to 50°C. The experiment results have shown remarkably high temperature sensitivities of 0.616nm/°C for 18cm and 0.31nm/°C for 40cm long cavity filter, respectively. The 18cm long cavity filter has been subjected to strain variations up to around 550μ ε and the filter has exhibited strain sensitivities of 0.02499nm/μ ε and 0.012nm/μ ε for two straining situations, where its cavity middle section of 18cm and 9cm were stretched, respectively. © 2012 SPIE.

Simultaneous refractive index and temperature measurement using cascaded long-period grating in double-cladding fibre

A compact scheme for simultaneous temperature and surrounding refractive index (SRI) measurement using two long-period gratings (LPGs) of different periods inscribed side-by-side in a single piece of a double-cladding fibre is presented. One of the LPGs is sensitive to both SRI and temperature, whilst the second is sensitive to temperature only.

The variant rpoS allele of S. enteritidis strain 27655R does not affect virulence in a chick model nor constitutive curliation but does generate a cold-sensitive phenotype

Adherence of pathogenic Escherichia coli and Salmonella spp. to host cells is in part mediated by curli fimbriae which, along with other virulence determinants, are positively regulated by RpoS. Interested in the role and regulation of curli (SEF17) fimbriae of Salmonella enteritidis in poultry infection, we tested the virulence of naturally occurring S. enteritidis PT4 strains 27655R and 27655S which displayed constitutive and null expression of curli (SEF17) fimbriae, respectively, in a chick invasion assay and analysed their rpoS alleles. Both strains were shown to be equally invasive and as invasive as a wild-type phage type 4 strain and an isogenic derivative defective for the elaboration of curli. We showed that the rpoS allele of 27655S was intact even though this strain was non-curliated and we confirmed that a S. enteritidis rpoS::strr null mutant was unable to express curli, as anticipated. Strain 27655R, constitutively curliated, possessed a frameshift mutation at position 697 of the rpoS coding sequence which resulted in a truncated product and remained curliated even when transduced to rpoS::strr. Additionally, rpoS mutants are known to be cold-sensitive, a phenotype confirmed for strain 27655R. Collectively, these data indicated that curliation was not a significant factor for pathogenesis of S. enteritidis in this model and that curliation of strains 27655R and 27655S was independent of RpoS. Significantly, strain 27655R possessed a defective rpoS allele and remained virulent. Here was evidence that supported the concept that different naturally occurring rpoS alleles may generate varying virulence phenotypic traits.

Simultaneous refractive index and temperature measurement using a cascaded long-period grating device

We present a compact scheme for simultaneous temperature and surrounding refractive index (SRI) measurement using two long period gratings (LPGs) of different periods inscribed side-by-side in a single piece of a double-cladding fibre. One of the LPGs is sensitive to both SRI and temperature changes whilst the second is SRI-insensitive but shows spectral shift with temperature changes. In addition, we show that a resonance peak of the SRI-insensitive LPG can be designed to appear in the EDFA wavelength region with potential use for gain flattening applications.

The Single drop drying of heat-sensitive materials

The literature relating to the principles and practice of drying of materials, particularly those susceptible to thermal degradation or undesirable loss of volatile components, has been reviewed. Single droplets of heat-sensitive materials were dried whilst suspended in a horizontal wind tunnel from a specially-designed, rotating thermocouple which enabled direct observation of drying behaviour and continuous measurement of droplet temperature as drying progressed. The effects of drying air temperature and initial solids concentration on the potency of various antibiotics, viz. ampicillin, chloramphenicol, oxytetracycline, streptomycin and tetracycline, were assessed using a modified Drug Sensitivity Testing technique. Only ampicillin was heat-sensitive at temperatures above 100°C, e.g. at an air temperature of 115°C its zone diameter was reduced from 100% to 45%. Selected enzymes, viz. dextran sucrase and invertase, were also dried and their residual activities determined by High Performance Liquid Chromatography. The residual activity of dextran sucrase was rapidly reduced at temperatures above 65°C, and the residual activity of invertase reduced rapidly at temperatures above 65°C; but drying with short residence times will retain most of its activity. The performance of various skin-forming encapsulants, viz. rice and wheat starch, dextrin, coffee, skim milk, fructose, gelatine 60 and 150 Bloom, and gum arabic, was evaluated to determine their capabilities for retention of ethanol as a model volatile, under different operating conditions. The effects of initial solids concentration, air velocity and temperature were monitored for each material tested. Ethanol content was analysed by Gas Liquid Chromatography and in some cases dried crusts were removed for examination. Volatiles retention was concluded to depend in all cases upon the rate and nature of the skin formation and selective diffusion phenomena. The results provided further insight into the inter-relationship between temperature, residence time and thermal degradation of heat-sensitive materials. They should also assist in selection of the preferred dryer for such materials, and of the operating parameter to enable maximum retention of the required physico-chemical characteristics in the dried materials.

Simultaneous measurement of temperature and strain: cross sensitivity considerations

Interferometric sensors using optical fibers as a transduction medium have been shown to be sensitive to a variety of physical measurands. A result of this is that the resolution of a system designed to sense strain, for example, may be compromised by fluctuations in the temperature of the environment. The possibility of simultaneously determining the strain and temperature applied to the same piece of highly birefringent fiber is discussed. Second-order effects are shown to be important for long sensing lengths or in the presence of high strains or temperature changes. The results of experiments carried out to verify the theoretical predictions are also described.

High temperature gas cleaning for advanced syngas utilization:Paper # 95

Biofuels and chemicals from biomass mean the gasification of biogenic feedstocks and the synthesis via methanol, dimethylester (DME) or Fischer-Tropsch products. To prevent the sensitive synthesis catalysts from poisoning the syngas must be free of tar and particulates. The trace concentrations of S-, C1-, N-species, alkali and heavy metals must be of the order of a few ppb. Moreover maximum conversion efficiency will be achieved performing the gas cleaning above the synthesis conditions. The concept of an innovative dry HTHP syngas cleaning is presented. Based on the HT particle filtration and suitable sorption and catalysis processes for the relevant contaminants a total concept will be derived, which leads to a syngas quality required for synthesis catalysts in only 2 combined stages. The experimental setup for the HT gas cleaning behind the 60 kWtherm entrained flow gasifier REGA of the institute is described. Results from HT filter experiments in pilot scale are presented. The performance of 2 natural minerals for HC1 and H2S sorption is discussed with respect to the parameters temperature, surface and residence time. Results from lab scale investigations on low temperature tar catalysts' performance (commercial and proprietary development) are discussed finally.

Fiber laser incorporating an intracavity micro-channel for refractive index and temperature sensing

In this letter, we present a standard linear cavity fiber laser incorporating a microchannel for refractive index (RI) and temperature sensing. The microchannel of ~6µm width created by femtosecond laser aided chemical etching provides an access to the external liquid; therefore, the laser cavity loss changes with the liquids of different RIs. Thus, at a fixed pump power, the output laser power will vary with the change of RI in the microchannel. The results show that the proposed sensing system has a linear response to both the surrounding medium RI and temperature. The RI sensitivity of the laser system is on the order of 10-3, while the temperature sensitivity is about 0.02 C. Both sensitivities could be further enhanced by employing a more sensitive photodetector and using higher pump power.

Simultaneous refractive index and temperature measurement using cascaded long-period grating in double-cladding fibre

A compact scheme for simultaneous temperature and surrounding refractive index (SRI) measurement using two long-period gratings (LPGs) of different periods inscribed side-by-side in a single piece of a double-cladding fibre is presented. One of the LPGs is sensitive to both SRI and temperature, whilst the second is sensitive to temperature only.