Switching the mode of sucrose utilization by Saccharomyces cerevisiae

Abstract Background Overflow metabolism is an undesirable characteristic of aerobic cultures of Saccharomyces cerevisiae during biomass-directed processes. It results from elevated sugar consumption rates that cause a high substrate conversion to ethanol and other bi-products, severely affecting cell physiology, bioprocess performance, and biomass yields. Fed-batch culture, where sucrose consumption rates are controlled by the external addition of sugar aiming at its low concentrations in the fermentor, is the classical bioprocessing alternative to prevent sugar fermentation by yeasts. However, fed-batch fermentations present drawbacks that could be overcome by simpler batch cultures at relatively high (e.g. 20 g/L) initial sugar concentrations. In this study, a S. cerevisiae strain lacking invertase activity was engineered to transport sucrose into the cells through a low-affinity and low-capacity sucrose-H+ symport activity, and the growth kinetics and biomass yields on sucrose analyzed using simple batch cultures. Results We have deleted from the genome of a S. cerevisiae strain lacking invertase the high-affinity sucrose-H+ symporter encoded by the AGT1 gene. This strain could still grow efficiently on sucrose due to a low-affinity and low-capacity sucrose-H+ symport activity mediated by the MALx1 maltose permeases, and its further intracellular hydrolysis by cytoplasmic maltases. Although sucrose consumption by this engineered yeast strain was slower than with the parental yeast strain, the cells grew efficiently on sucrose due to an increased respiration of the carbon source. Consequently, this engineered yeast strain produced less ethanol and 1.5 to 2 times more biomass when cultivated in simple batch mode using 20 g/L sucrose as the carbon source. Conclusion Higher cell densities during batch cultures on 20 g/L sucrose were achieved by using a S. cerevisiae strain engineered in the sucrose uptake system. Such result was accomplished by effectively reducing sucrose uptake by the yeast cells, avoiding overflow metabolism, with the concomitant reduction in ethanol production. The use of this modified yeast strain in simpler batch culture mode can be a viable option to more complicated traditional sucrose-limited fed-batch cultures for biomass-directed processes of S. cerevisiae.

The relationship between the speech perception and the degree of satisfaction among adult users of cochlear implants

Introduction: In recent years, the benefits associated with the use of cochlear implants (CIs), especially with regard to speech perception, have proven to surpass those produced by the use of hearing aids, making CIs a highly efficient resource for patients with severe/profound hearing loss. However, few studies so far have assessed the satisfaction of adult users of CIs. Objective: To analyze the relationship between the level of speech perception and degree of satisfaction of adult users of CI. Method: This was a prospective cross-sectional study conducted in the Audiological Research Center (CPA) of the Hospital of Craniofacial Anomalies, University of São Paulo (HRAC/USP), in Bauru, São Paulo, Brazil. A total of 12 users of CIs with pre-lingual or post-lingual hearing loss participated in this study. The following tools were used in the assessment: a questionnaire, "Satisfaction with Amplification in Daily Life" (SADL), culturally adapted to Brazilian Portuguese, as well as its relationship with the speech perception results; a speech perception test under quiet conditions; and the Hearing in Noise Test (HINT)Brazil under free field conditions. Results: The participants in the study were on the whole satisfied with their devices, and the degree of satisfaction correlated positively with the ability to perceive monosyllabic words under quiet conditions. The satisfaction did not correlate with the level of speech perception in noisy environments. Conclusion: Assessments of satisfaction may help professionals to predict what other factors, in addition to speech perception, may contribute to the satisfaction of CI users in order to reorganize the intervention process to improve the users' quality of life.

Influence of miniscrew dental root proximity on its degree of late stability

Purpose of this study was to evaluate the influence of miniscrew dental root proximity on its late stability degree. 40 miniscrews were inserted between maxillary second premolars and first molars for anterior retraction. Post-surgical radiographs were used to measure the septum width in the insertion site, insertion height, distal and mesial distance from miniscrew to dental root, and the smallest distance between miniscrew and dental root. The miniscrews were divided in two groups according to septum width: ?3 mm (20 miniscrews) and >3 mm (20 miniscrews). The soft tissue in the insertion site, sensitivity during load, plaque around the miniscrew, and evaluation period were also considered. The results showed no significant difference in miniscrew mobility degree and success rate between groups. Miniscrew dental root proximity did not influence the stability and success rate when the distance between the miniscrew and dental root indicated no periodontal ligament invasion. The overall success rate was 90% and no variable was associated with miniscrew failure. Nevertheless, patient sensitivity was frequently associated with some degree of mini-implant mobility. The septum width did not influence the stability and success rate of this anchorage system, but the extreme root proximity did.

Degree of satisfaction among hearing aid users

Introduction: Hearing loss (HL) is defined as the complete or partial loss of hearing ability. Aims: To characterize (1) the degree of satisfaction among adult and elderly hearing aid (HA) users who were treated by a public hearing health service and (2) the relationship between satisfaction and the variables of gender, age, degree of HL, and type of HA. Method: The clinical and experimental study included the administration of the Satisfaction with Amplification in Daily Life (SADL) questionnaire to 110 patients who had used HAs for more than 3 months and were 18 years of age or older. Results: Test patients were sex-balanced (48% were women) and had a mean age of 67 years. A relatively high incidence of sensorineural moderate HL was detected in the study patients (66%) and device B was the most commonly used HA type (48%). No significant differences were evident between HA satisfaction and sex. The importance placed on services/costs and personal image varied between age groups. Correlation was evident at all levels between user satisfaction and amplification. Decreased satisfaction was observed in individuals with severe and/or profound HL. The type of HA used yielded statistically significant differences in the positive effects referring. Conclusion: No correlations were evident between the different factors proposed. HA users exhibited high levels of satisfaction in all SADL areas

Degree of multicollinearity and variables involved in linear dependence in additive-dominant models

The objective of this work was to assess the degree of multicollinearity and to identify the variables involved in linear dependence relations in additive-dominant models. Data of birth weight (n=141,567), yearling weight (n=58,124), and scrotal circumference (n=20,371) of Montana Tropical composite cattle were used. Diagnosis of multicollinearity was based on the variance inflation factor (VIF) and on the evaluation of the condition indexes and eigenvalues from the correlation matrix among explanatory variables. The first model studied (RM) included the fixed effect of dam age class at calving and the covariates associated to the direct and maternal additive and non-additive effects. The second model (R) included all the effects of the RM model except the maternal additive effects. Multicollinearity was detected in both models for all traits considered, with VIF values of 1.03 - 70.20 for RM and 1.03 - 60.70 for R. Collinearity increased with the increase of variables in the model and the decrease in the number of observations, and it was classified as weak, with condition index values between 10.00 and 26.77. In general, the variables associated with additive and non-additive effects were involved in multicollinearity, partially due to the natural connection between these covariables as fractions of the biological types in breed composition.

Metabolism of plasma cholesterol and lipoprotein parameters are related to a higher degree of insulin sensitivity in high HDL-C healthy normal weight subjects

Abstract Background We have searched if plasma high density lipoprotein-cholesterol (HDL-C) concentration interferes simultaneously with whole-body cholesterol metabolism and insulin sensitivity in normal weight healthy adult subjects. Methods We have measured the activities of several plasma components that are critically influenced by insulin and that control lipoprotein metabolism in subjects with low and high HDL-C concentrations. These parameters included cholesteryl ester transfer protein (CETP), phospholipid transfer protein (PLTP), lecithin cholesterol acyl transferase (LCAT), post-heparin lipoprotein lipase (LPL), hepatic lipase (HL), pre-beta-1HDL, and plasma sterol markers of cholesterol synthesis and intestinal absorption. Results In the high-HDL-C group, we found lower plasma concentrations of triglycerides, alanine aminotransferase, insulin, HOMA-IR index, activities of LCAT and HL compared with the low HDL-C group; additionally, we found higher activity of LPL and pre-beta-1HDL concentration in the high-HDL-C group. There were no differences in the plasma CETP and PLTP activities. Conclusions These findings indicate that in healthy hyperalphalipoproteinemia subjects, several parameters that control the metabolism of plasma cholesterol and lipoproteins are related to a higher degree of insulin sensitivity.

Two-degree-of-freedom vortex-induced vibration of circular cylinders with very low aspect ratio and small mass ratio

The investigation of vortex-induced vibration on very short cylinders with two degrees of freedom has drawn the attention of a large number of researchers. Some investigations on such a problem are carried out in order to have a better understanding of the physics involved in vortex-induced motions of floating bodies such as offshore platforms. In this paper, experiments were carried out in a recirculating water channel over the range of Reynolds number 6000degrees of freedom, three different small mass ratios (m⁎=1.00; 2.62 and 4.36) and very low aspect ratios (0.3≤L/D≤2.0) were shown and the results were discussed in depth. Conversely to what would be expected for cylinders with very low aspect ratio, the results showed large motions in the transverse direction with maximum amplitudes around 1.5 diameters for cylinders with L/D=2.0, despite being smaller when the aspect ratio is reduced. Moreover, the response amplitudes presented high values around 0.4 diameters in the in-line direction. In fact, the large transverse motions were related to a strong coupling with the in-line responses, visibly identified in the plots of nondimensional frequency, as well as by the trajectories in the XY-plane, Lissajous figures, particularly in the case of m⁎=1.00 and L/D=2.0, when 8-shape trajectories were clearly observed. The case of m⁎=1.00 deserves more attention because of its smaller amplitude compared to the cases with the same aspect ratio and a larger mass ratio. This counter-intuitive behavior seems to be related to the energy transferring process from the steady stream to the oscillatory hydroelastic system. Finally, it is noteworthy that the characteristic of the “Strouhal-like” number decreases when the aspect ratio decreases, as also observed in previous works available in the literature, most of them for stationary cylinders.

Two-degree-of-freedom vortex-induced vibration of circular cylinders with very low aspect ratio and small mass ratio

The investigation of vortex-induced vibration on very short cylinders with two degrees of freedom has drawn the attention of a large number of researchers. Some investigations on such a problem are carried out in order to have a better understanding of the physics involved in vortex-induced motions of floating bodies such as offshore platforms. In this paper, experiments were carried out in a recirculating water channel over the range of Reynolds number 6000degrees of freedom, three different small mass ratios (m⁎=1.00; 2.62 and 4.36) and very low aspect ratios (0.3≤L/D≤2.0) were shown and the results were discussed in depth. Conversely to what would be expected for cylinders with very low aspect ratio, the results showed large motions in the transverse direction with maximum amplitudes around 1.5 diameters for cylinders with L/D=2.0, despite being smaller when the aspect ratio is reduced. Moreover, the response amplitudes presented high values around 0.4 diameters in the in-line direction. In fact, the large transverse motions were related to a strong coupling with the in-line responses, visibly identified in the plots of nondimensional frequency, as well as by the trajectories in the XY-plane, Lissajous figures, particularly in the case of m⁎=1.00 and L/D=2.0, when 8-shape trajectories were clearly observed. The case of m⁎=1.00 deserves more attention because of its smaller amplitude compared to the cases with the same aspect ratio and a larger mass ratio. This counter-intuitive behavior seems to be related to the energy transferring process from the steady stream to the oscillatory hydroelastic system. Finally, it is noteworthy that the characteristic of the “Strouhal-like” number decreases when the aspect ratio decreases, as also observed in previous works available in the literature, most of them for stationary cylinders.

Utilization of fringe projection technique for evaluation of wound dimensions and of healing progress.

Background: The methods used for evaluating wound dimensions, especially the chronic ones, are invasive and inaccurate. The fringe projection technique with phase shift is a non-invasive, accurate and low-cost optical method. Objective: The aim is to validate the technique through the determination of dimensions of objects of known topography and with different geometries and colors to simulate the wounds and tones of skin color. Taking into account the influence of skin wound optical factors, the technique will be used to evaluate actual patients’ wound dimensions and to study its limitations in this application. Methods: Four sinusoidal fringe patterns, displaced ¼ of period each, were projected onto the objects surface. The object dimensions were obtained from the unwrapped phase map through the observation of the fringe deformations caused by the object topography and using phase shift analysis. An object with simple geometry was used for dimensional calibration and the topographic dimensions of the others were determined from it. After observing the compatibility with the data and validating the method, it was used for measuring the dimensions of real patients’ wounds. Results and Conclusions: The discrepancies between actual topography and dimensions determined with Fringe Projection Technique and for the known object were lower than 0.50 cm. The method was successful in obtaining the topography of real patient’s wounds. Objects and wounds with sharp topographies or causing shadow or reflection are difficult to be evaluated with this technique.

Determination of degree of ionization of poly(allylamine hydrochloride) (PAH) and poly[1-[4-(3-carboxy‑4 hydroxyphenylazo)benzene sulfonamido]-1,2-ethanediyl, sodium salt] (PAZO) in layer-by-layer films using vacuum photoabsorption spectroscopy

Electrostatic and hydrophobic interactions govern most of the properties of supramolecular systems, which is the reason determining the degree of ionization of macromolecules has become crucial for many applications. In this paper, we show that highresolution ultraviolet spectroscopy (VUV) can be used to determine the degree of ionization and its effect on the electronic excitation energies of layer-by-layer (LbL) films of poly(allylamine hydrochloride) (PAH) and poly[1-[4-(3-carboxy-4 hydroxyphenylazo)- benzene sulfonamido]-1,2-ethanediyl, sodium salt] (PAZO). A full assignment of the VUV peaks of these polyelectrolytes in solution and in cast or LbL films could be made, with their pH dependence allowing us to determine the p'K IND. a' using the Henderson-Hasselbach equation. The p'K IND. a' for PAZO increased from ca. 6 in solution to ca. 7.3 in LbL films owing to the charge transfer from PAH. Significantly, even using solutions at a fixed pH for PAH, the amount adsorbed on the LbL films still varied with the pH of the PAZO solutions due to these molecular-level interactions. Therefore, the procedure based on a comparison of VUV spectra from solutions and films obtained under distinct conditions is useful to determine the degree of dissociation of macromolecules, in addition to permitting interrogation of interface effects in multilayer films.

Evaluation of degree of conversion and hardness of dental composites photo-activated with different light guide tips

Objective: The aim of this study was to evaluate the degree of conversion and hardness of different composite resins, photo-activated for 40 s with two different light guide tips, fiber optic and polymer. Methods: Five specimens were made for each group evaluated. The percentage of unreacted carbon double bonds (% C=C) was determined from the ratio of absorbance intensities of aliphatic C=C (peak at 1637 cm-1) against internal standard before and after curing of the specimen: aromatic C-C (peak at 1610 cm-1). The Vickers hardness measurements were performed in a universal testing machine. A 50 gf load was used and the indenter with a dwell time of 30 seconds. The degree of conversion and hardness mean values were analyzed separately by ANOVA and Tukey's test, with a significance level set at 5%. Results: The mean values of degree of conversion for the polymer and fiber optic light guide tip were statistically different (P<.001). The hardness mean values were statistically different among the light guide tips (P<.001), but also there was difference between top and bottom surfaces (P<.001). Conclusions: The results showed that the resins photo-activated with the fiber optic light guide tip promoted higher values for degree of conversion and hardness.

Similar carbohydrate but enhanced lactate utilization during exercise after 9 wk of acclimatization to 5,620 m

[EN] We hypothesized that reliance on lactate as a means of energy distribution is higher after a prolonged period of acclimatization (9 wk) than it is at sea level due to a higher lactate Ra and disposal from active skeletal muscle. To evaluate this hypothesis, six Danish lowlanders (25 +/- 2 yr) were studied at rest and during 20 min of bicycle exercise at 146 W at sea level (SL) and after 9 wk of acclimatization to 5,260 m (Alt). Whole body glucose Ra was similar at SL and Alt at rest and during exercise. Lactate Ra was also similar for the two conditions at rest; however, during exercise, lactate Ra was substantially lower at SL (65 micro mol. min(-1). kg body wt(-1)) than it was at Alt (150 micro mol. min(-1). kg body wt(-1)) at the same exercise intensity. During exercise, net lactate release was approximately 6-fold at Alt compared with SL, and related to this, tracer-calculated leg lactate uptake and release were both 3- or 4-fold higher at Alt compared with SL. The contribution of the two legs to glucose disposal was similar at SL and Alt; however, the contribution of the two legs to lactate Ra was significantly lower at rest and during exercise at SL (27 and 81%) than it was at Alt (45 and 123%). In conclusion, at rest and during exercise at the same absolute workload, CHO and blood glucose utilization were similar at SL and at Alt. Leg net lactate release was severalfold higher, and the contribution of leg lactate release to whole body lactate Ra was higher at Alt compared with SL. During exercise, the relative contribution of lactate oxidation to whole body CHO oxidation was substantially higher at Alt compared with SL as a result of increased uptake and subsequent oxidation of lactate by the active skeletal muscles.