Fluorescence enhancement from plasmonic Au templates

We have fabricated sub-quarter-micron-patterned Au templates with electron beam lithography, and studied their effect on the fluorescence intensity of immobilized, anti-rabbit IgG antibody labeled with AlexaFluor® 546. Varying the geometry of the structured surface, the plasmon resonances are tuned to match the fluorescence wavelengths and achieve significant fluorescence enhancement. Full electrodynamic simulations were used to understand the optical response and access the quality of the fabricated structures for surface plasmon excitation.     

Biosynthesis of lipids and organic acids by Yarrowia lipolytica strains cultivated on glucose

The biochemical behavior of wild‐type or genetically modified (presenting decreased expression of intracellular acyl‐CoA oxidases) Yarrowia lipolytica strains cultivated on commercial glucose was studied. Flask nitrogen‐limited cultures were performed favoring the production of organic acids (and potentially the accumulation of lipid). Nitrogen depletion induced secretion of citric acid, while intracellular lipid was not produced in high quantities. Maximum total citric acid up to 49 g/L (yield 0.85 g/g glucose) was produced. In some of the wild‐type strains, an increase of glucose in the medium also induced noticeable production of acetic acid. Increasing the amount of added glucose led to an increase in the total lipid quantity (%) produced, although in the stationary growth phase the concentration of lipid declined, indicating lipid degradation even for the genetically modified strains. Total lipid amount did not exceed the value of 14%, while neutral fractions increased with increase in glucose concentration. In all cases, the total microbial lipids and major lipid fractions were composed of C₁₆ and C₁₈ (principally unsaturated) fatty acids. Finally, in several of the strains cultured in media containing a low glucose concentration, unicellular morphology was observed, while at high glucose concentrations mycelia were predominant.     

On the impact of urban climate on the energy consumption of buildings

Climatic measurements from almost 30 urban and suburban stations as well as specific measurements performed in 10 urban canyons in Athens, Greece, have been used to assess the impact of the urban climate on the energy consumption of buildings. It is found that for the city of Athens, where the mean heat island intensity exceeds 10°C, the cooling load of urban buildings may be doubled, the peak electricity load for cooling purposes may be tripled especially for higher set point temperatures, while the minimum COP value of air conditioners may be decreased up to 25% because of the higher ambient temperatures. During the winter period, the heating load of centrally located urban buildings is found to be reduced up to 30%. Regarding the potential of natural ventilation techniques when applied to buildings located in urban canyons, it is found that, mainly during the day, this is seriously reduced because of the important decrease of the wind speed inside the canyon. Air flow reduction may be up to 10 times the flow that corresponds to undisturbed ambient wind conditions.     

Active voltage clamp in flyback converters operating in CCM mode under wide load variation

Active clamp topologies of low power dissipation have become a very attractive solution in order to limit overvoltages in flyback converters. Although many suitable topologies have been introduced for the case of discontinuous conduction mode (DCM), where the duty cycle value depends on the load level, in continuous conduction mode (CCM) it is more difficult to appropriately design such topologies so as to "sense" load changes-due to the small duty cycle divergence under wide load variation. Taking for granted that in order to achieve high power-factor correction in these converters, CCM is a more attractive mode of operation, a drastic solution for this case that will manage to eliminate voltage stresses under wide load changes has become very essential. For this purpose, this paper presents an active clamp topology with small power dissipation, suitable for flyback converters operating in CCM mode. Its main idea is the use of a load-dependent current source, consisting of an auxiliary converter operating in DCM mode. Experimental results highlight the effectiveness of the proposed topology under wide load changes, establishing it as an appropriate solution in order to develop flyback converters, even at the power range of 500 W.     

Screening various Yarrowia lipolytica strains for citric acid production

Citric acid (CA) productivity by Yarrowia lipolytica dependents on strain type, carbon source, carbon to nitrogen (C/N) molar ratio as well as physicochemical conditions (pH, temperature, oxygen transfer rate, etc.). In the current study, 10 different Y. lipolytica strains were first challenged in shake-flask culture for CA production in a glucose-based media under nitrogen-limited conditions. For the most promising one, strain K57, CA productivity was monitored during culture in batch bioreactor for three initial C/N molar ratio (167, 367, and 567 Cmol/Nmol). The highest CA yield (0.77 g/g glucose), titre (72.3 g/L CA), and productivity (0.04 g/g.h) were found for C/N ratio of 367. However, the highest growth rate was obtained for an initial C/N ratio of 167. From these results, Y. lipolytica strain K57 could be considered to produce CA at higher titre on glucose-based medium in batch bioreactor than others Y. lipolytica strain reported in the literature.     

Effects of chromium (VI) addition on the activated sludge process

The effect of hexavalent chromium, Cr(VI), addition on various operating parameters of activated sludge process was evaluated. To accomplish this, two parallel lab-scale continuous-flow activated sludge plants were operated. One was used as a control plant, while the other received Cr(VI) concentrations equal to 0.5, 1, 3 and 5 mgl(-1). Cr(VI) concentrations of 0.5 mgl(-1) caused significant inhibition of the nitrification process (up to 74% decrease in ammonia removal efficiency). On the contrary, the effect of Cr(VI) on organic substrate removal was minor for concentrations up to 5 mgl(-1), indicating that heterotrophic microorganisms are less sensitive to Cr(VI) than nitrifiers. Activated sludge floc size and structure characterization showed that Cr(VI) concentrations higher than 1 mgl(-1) reduced the filaments abundance, causing the appearance of pin-point flocs and free-dispersed bacteria. Additionally, the variability of protozoa and rotifers was reduced. As a result of disperse growth, effluent quality deteriorated, since significant amounts of suspended solids escaped with the effluent. Termination of Cr(VI) addition led to a partial recovery of the nitrification process (up to 57% recovery). Similar recovery signs were not observed for activated sludge floc size and structure. Finally, shock loading to the control plant with 5 mgl(-1) Cr(VI) for 2 days resulted in a significant inhibition of the nitrification process and a reduction in filamentous microorganisms abundance.     

Expression of retinoblastoma gene product and p21 (WAF1/Cip 1) protein in gliomas: correlations with proliferation markers, p53 expression and survival

AIMS/BACKGROUND: The construction and validation of an instrument for the assessment of subjective visual disability in the cataract patient is described. This instrument is specifically designed for measuring the outcome of cataract surgery with respect to visual disability. METHODS: Visually related activities thought to be affected by cataract were considered for the questionnaire. These were reduced by pilot study and principal components analysis to 18 items. A patient's assessment of his/her ability to perform each task was scored on a four point scale. Scores were averaged to create an overall index of visual disability, as well as subscale indices for mobility related disability, distance/lighting/reading related disability, and near and related tasks visual disability. The questionnaire, administered verbally is entitled "The Visual Disability Assessment (VDA)". Reliability testing included test-retest reliability, interobserver reliability (p, the intraclass correlation coefficient), and internal consistency reliability (Cronbach's alpha). Construct validation, the process for proving that a test measures what it is supposed to measure, included consideration of content validity, comparison with the established Activities of Daily Vision Scale (ADVS) and empirical support with factor analysis. RESULTS: For the four indices, interobserver reliability varied from 0.92 to 0.94, test-retest reliability varied from 0.96 to 0.98, and internal consistency reliability varied from 0.80 to 0.93. The VDA compared favourably with the ADVS by correlation, but Bland-Altman analysis demonstrated that the two instruments were not clinically interchangeable. Factor analysis suggests that all test items measure a common theme, and the subgroupings reflect common themes. CONCLUSIONS: The VDA is easy to administer because it has a short test time and scoring is straightforward. It has excellent interobserver, test-retest, and internal consistency reliability, and compares favourably with the ADVS, another test of visual disability. Factor analysis demonstrated that the 18 items measure a related theme, which can be assumed to be visual disability. The VDA is a valid instrument which provides a comprehensive assessment of visual disability in cataract patients and is designed to detect changes within a patient over time.     

Lipids of oleaginous yeasts. Part II: Technology and potential applications

The process of lipid accumulation in the oleaginous yeasts cultivated in various fermentation configurations when either sugars and related compounds or hydrophobic substances are used as substrates is presented and kinetic models describing both de novo and ex novo lipid accumulation are analyzed. Technological aspects related with single cell oil (SCO) produced by oleaginous yeasts are depicted. The influence of culture parameters upon lipid production process is presented. Lipid production has been studied in batch, fed‐batch, and continuous cultivation systems using yeasts belonging to the species Lipomyces starkeyi, Rhodosporidium toruloides, Apiotrichum curvatum, Candida curvata, Cryptococcus curvatus, Trichosporon fermentans, and Yarrowia lipolytica. The potentiality of yeasts to produce SCO as starting material of 2nd generation biodiesel is indicated and discussed. Of significant importance is also the utilization of yeast lipids as substitutes of high added value exotic fats (e.g., cocoa butter). Lipid produced by the various yeasts presents, in general, similar composition with that of common vegetable oils being composed of unsaturated fatty acids, whereas cocoa butter is principally composed of saturated fatty acids, consequently the various strategies that are followed in order to increase the cellular saturated fatty acid content of the yeast lipid are presented and comprehensively discussed.     

Substrate Specificity of the Highly Thermostable Esterase EstDZ3

Esterases are among the most studied enzymes, and their applications expand into several branches of industrial biotechnology. Yet, despite the fact that information on their substrate specificity is crucial for selecting or designing the best fitted biocatalyst for the desired application, it cannot be predicted from their amino acid sequence. In this work, we studied the substrate scope of the newly discovered hydrolytic extremozyme, EstDZ3, against a library of esters with variable carbon chain lengths in an effort to understand the crucial amino acids for the substrate selectivity of this enzyme. EstDZ3 appears to be active against a wide range of esters with high selectivity towards medium- to long-carbon chain vinyl esters. In-silico studies of its 3D structure revealed that the selectivity might arise from the mainly hydrophobic nature of the active site's environment.