Increased number of cardiomyocytes in cross-sections from tachycardia-induced cardiomyopathic hearts

Based on positive identification of DNA replication and mitotic division in cardiomyocytes isolated from failing hearts, it has been proposed that adult ventricular cardiomyocytes can gain the capacity to proliferate with progression of heart failure. However, due to the lack of a reliable method to distinctly image individual cardiac cells within the myocardial syntitium, such a concept still remains largely controversial. In the present study, we used laser confocal microscopy, to image cross-sections of intact myocardium stained with fluorescein-conjugated wheat germ agglutinin and propidium iodide. This approach allowed to clearly separate the profile of individual myocytes within cardiac tissue sections. We found that in the left ventricles of dogs, subjected to tachycardia-induced cardiomyopathy, the number of cells was significantly increased in both longitudinal and transversal sections. Treatment with the angiotensin-converting enzyme inhibitor, enalapril, reversed these changes to values similar to those found in controls. Therefore, this study provides evidence, at the in situ level, for cellular hyperplasia in heart failure. This supports the more general notion that adult cardiomyocytes may not be terminally differentiated, and that an increase in cell number could contribute to the increase in left ventricular mass observed with progression of disease.     

Metabolic oxidative stress activates signal transduction and gene expression during glucose deprivation in human tumor cells

The mechanism of glucose deprivation-induced activation of Lyn kinase (Lyn), c-Jun N-terminal kinase 1 (JNK1) and increased expression of basic fibroblast growth factor (bFGF) and c-Myc was investigated in MCF-7/ADR adriamycin-resistant human breast carcinoma cells. Glucose deprivation significantly increased steady state levels of oxidized glutathione content (GSSG) and intracellular prooxidants (presumably hydroperoxides) as well as caused the activation of Lyn, JNK1, and the accumulation of bFGF and c-Myc mRNA. The suppression of GSSG accumulation and prooxidant production by treatment with the thiol antioxidant, N-acetylcysteine, also suppressed all the increases in kinase activation and gene expression observed during glucose deprivation. In addition, glucose deprivation was shown to induce oxidative stress in IMR90 SV40 transformed human fibroblasts, indicating that this phenomena is not limited to the MCF-7/ADR cell line. These and previous observations from our laboratory show that glucose deprivation-induced oxidative stress in MCF-7/ADR cells activates signal transduction involving Lyn, JNK1, and mitogen activated protein kinases (ERK1/ERK2) which results in increased bFGF and c-Myc mRNA accumulation. These results provide support for the hypothesis that alterations in intracellular oxidation/reduction reactions link changes in glycolytic metabolism to signal transduction and gene expression in these human tumor cells.     

Robust control design of an activated sludge process

A H-inf control strategy is presented for a robustly performing activated sludge process. In operational terms, the objective is to conduct the process imposing that the biomass concentration in the recycle stream follows closely a time-varying, process-dependent, reference signal. The corresponding control objective is described as the development of a robust reference-tracking control structure with the best possible disturbance compensation, able to perform with noisy measurements and able to cope with variations in key process model parameters. The proposed algorithm embeds an estimation of states by solving the Riccati equation and avoids parameter estimation by assuming their variability within known bounds. Results are presented which show a favourable behaviour of the robust controller in comparison with a conventional PI control structure. Copyright © 1999 John Wiley & Sons, Ltd.     

Stability and stabilisation of penicillin acylase

The stability of an oligomeric enzyme, penicillin acylase, was studied in aqueous media. The enzyme was produced by mutant cells of Escherichia coli ATCC 9637, extracted from the periplasmic space by osmotic shock and further purified using a pseudo‐affinity adsorption process. Enzyme stabilisation attempts were performed with salts, alcohols and sugars. The highest levels of retained activity were obtained in the presence of 15% (w/v) ammonium or sodium sulfate. A kinetic model was proposed to describe the inactivation of penicillin acylase, taking into account results obtained in stability assays performed at different temperatures and with different enzyme concentrations. According to this model, the inactivation of penicillin acylase involves an intermediary active precursor of the enzyme, formed prior to dissociation into sub‐units. © 1999 Society of Chemical Industry     

Microstructural Development of an AlNiBi Alloy and Influence of the Transient Horizontal Solidification Parameters on Microhardness

The combination of the structural and tribological properties presented by AlNiBi alloys has motivated us to establish, as the main objective of this study, the investigation of the microstructural evolution and its influence on the microhardness (HV) of Al-3wt pct Ni-1wt pct Bi alloy horizontally solidified via a water-cooled directional solidification device. Temperature mapping by thermocouples inserted in the metal has been performed for experimental determination of the solidification thermal parameters, such as the growth rate and cooling rate (V_L and T_R, respectively). The microstructure has been characterized by optical and scanning electron microscopy and by microanalysis of the composition via dispersive energy spectroscopy (EDS composition). The macrostructure of the as-solidified ingot is characterized by columnar grains, and the final microstructure consists of an Al-rich primary phase (α-Al) and a eutectic mixture composed of two phases: α-Al + Al₃Ni intermetallic (β) with Bi particles anchored on the β phase. The Bi droplet scale is affected by the thermal parameters. The primary phase (α-Al) is characterized by a reverse cellular-to-dendritic microstructural transition. Cellular and dendritic microstructures have been quantified by the cell, primary dendrite arm, secondary dendrite arm, and tertiary dendrite arm spacings (λ_C, λ₁, λ₂, and λ₃, respectively). The relationships of λ_C, λ₁, λ₂, and λ₃ with V_L and T_R have been established via power-type mathematical expressions. The HV dependence on λ_C, λ₁, λ₂, and λ₃ has been analyzed in both cellular and dendritic microstructural zones. It has been observed that the HV values do not vary in the dendritic zone; however, Hall–Petch’s mathematical equations characterize the HV variation with these thermal and microstructural parameters in the cellular zone.

     

Absorption of anthocyanins through intestinal epithelial cells – Putative involvement of GLUT2

Anthocyanins bioavailability is a major issue regarding their biological effects and remains unclear due to few data available on this matter. This work aimed to evaluate the absorption of anthocyanins at the intestine using Caco‐2 cells. Anthocyanin extract, rich in malvidin‐3‐glucoside, was obtained from red grape skins and tested on Caco‐2 cells. The absorption of anthocyanins, in absence or presence of 1% ethanol, was detected by HPLC/DAD/LC‐MS. Our results showed that this transport was significantly increased in the presence of ethanol especially after 60 min of incubation. In addition, cells that were pretreated for 96 h with anthocyanins (200 μg/mL) showed an increase of their own transport (about 50% increase). Expression of glucose transporters sodium‐dependent glucose transporter 1, facilitative glucose transporters 5, and facilitative glucose transporters 2 was assessed by RT‐PCR. It was found that facilitative glucose transporters 2 expression was increased (60%) in Caco‐2 cells pretreated with anthocyanins, by comparison with controls. When the effect of anthocyanin extract on ³H‐2‐deoxy‐D ‐glucose uptake was tested, an inhibitory effect was observed (about 60% decrease). However, the malvidin aglycone was tested and had no effect. In conclusion, anthocyanins could be absorbed through Caco‐2 cells, and can interfere with their own transport and also with glucose intestinal uptake.     

Effects of supplementing yeast culture to diets differing in starch content on performance and feeding behavior of dairy cows

The objectives were to evaluate the effects of a culture of Saccharomyces cerevisiae (YC) on lactation performance of cows fed diets differing in starch content. Fifty-six Holstein cows at 42 d postpartum were blocked by parity and milk production and randomly assigned to 1 of 4 treatments, low starch (23% diet DM) and no YC (LS-control), low starch and 15 g/d of YC (LS-YC), high starch (29% diet DM) and no YC (HS-control), and high starch and 15 g/d of YC (HS-YC). The experiment lasted 14 wk. Blood was sampled twice weekly during the first 5 wk in the experiment. Feeding behavior was evaluated in 2 consecutive days when cows were 33 d in the experiment. On d 92 in the experiment, cows were challenged with 3 kg of corn grain DM immediately before the morning feeding. Blood was sampled in the first 12 h after the challenge. Rumen fluid was collected 5 h after the challenge, and pH, ammonia N, short-chain fatty acids, and lactate concentrations were quantified. Lactation performance was measured daily before and after the challenge. Supplementation with YC increased yields of 3.5% fat-corrected milk and energy-corrected milk by 2.2 and 2.0 kg/d, and the increments were observed in both low- and high-starch diets. Feeding HS tended to decrease milk fat content (LS = 3.88 vs. HS = 3.73%), but increased concentration (LS = 2.87 vs. HS = 3.00%) and yield (LS = 1.11 vs. HS = 1.20 kg/d) of milk true protein. Feeding YC increased yields of fat and true protein in milk by 100 and 60 g/d. Energy balance, body weight, and feed efficiency did not differ with treatments. Feeding HS reduced eating time (LS = 177 vs. HS = 159 min/12 h) and intermeal interval (LS = 103 vs. HS = 82 min), but tended to increase eating rate (LS = 139 vs. HS = 150 g/min). Interactions were detected between level of starch and YC for ruminating time, meal duration, and meal size because within LS, feeding YC increased ruminating time 23 min/12 h, but reduced meal duration 6 min/meal and meal size 0.7 kg/meal. Concentrations of glucose in plasma increased (LS = 62.1 vs. HS = 63.8 mg/dL), whereas those of urea N decreased (LS = 10.1 vs. HS = 9.4 mg/dL) with feeding HS compared with LS in the first 5 wk in the experiment, and the same responses were observed after the challenge with corn grain. After the challenge, rumen pH was less and short-chain fatty acid concentrations were greater in cows fed HS compared with those fed LS; however, supplementing YC to high-starch diets increased rumen pH (HS-control = 5.72 vs. HS-YC = 6.12) and reduced concentrations of lactate in rumen fluid (HS-control = 7.72 vs. HS-YC = 1.33 mM) and haptoglobin in plasma 28%. Feeding YC improved lactation performance irrespective of the level of dietary starch and reduced the risk of subacute rumen acidosis induced by a grain challenge when cows were fed a high-starch ration.     

The Influence of the Rebaudioside A Content of Stevia (Stevia rebaudiana Bertoni) on the Determination of Sweetness Equivalence in Bittersweet Chocolates,Using the Time‐Intensity Analysis

The consumption of diet products has increased greatly in recent years. The objectives of the study were to develop a bittersweet chocolate added inulin and stevias with different rebaudioside A contents (60%, 80%, and 97%). Five chocolate samples were formulated with different sucrose concentrations to determine the ideal sucrose concentration for bittersweet chocolate. The use of just‐about‐right scale identified an ideal sucrose concentration of 47.5% (w/w). The sweetness equivalence in sugar‐free bittersweet chocolates was determined by the time–intensity method by 14 selected and trained judges. The data collected during each session of sensory evaluation furnished the following parameters in relation to the sweet stimulus: Imax (maximum intensity recorded), Timax (time at which the maximum intensity was recorded), Area (area of time × intensity curve), and Ttot (total duration time of the stimulus). The time–intensity analysis indicated that the percentages of rebaudioside A did not interfere with the sweetness intensity of the sweetener stevia in bittersweet chocolate and there was no significant difference in the concentrations tested (0.16%, 0.22%, 0.27%) of each stevia, in relation to the parameters evaluated. In addition, the reduction in fat content did not alter the perception of the sweetness intensity of the samples. These results showed important information to research and development of chocolate products. Therefore, the use of the lowest stevia concentration tested (0.16%) is the most indicated for use, since this quantity was sufficient to reach the ideal sweetness of the product, so there was no point in adding more.