Detection of cellular responses to toxicants by dielectrophoresis

The dielectrophoretic (DEP) crossover method has been applied to the detection of cell responses to toxicants. Time and dose responses of the human cultured leukemia (HL-60) line were measured for paraquat, styrene oxide (SO), N-nitroso-N-methylurea (NMU) and puromycin. These toxicants were chosen because of their different predominant mechanisms of action, namely membrane free radical attack, simultaneous membrane and nucleic acid attack, nucleic acid alkylation, and protein synthesis inhibition, respectively. For all treatments, the specific membrane capacitance (C_mem) of the cells decreased while the specific membrane conductance (G_mem) increased in dose- and time-dependent manners. The DEP responses correlated sensitively with alterations in cell surface morphology, especially folds, microvilli, and blebs, observed by scanning electron microscopy. The DEP method was more sensitive to agents that had a direct action on the membrane than to agents for which membrane alterations were secondary. The responses to paraquat and SO, which directly damaged the cell membrane, could be detected 15 min after exposure, while those for puromycin and NMU, which acted on intracellular targets, could be detected after 30 min. The detection times and dose sensitivity results showed that the DEP method is much faster and more sensitive than conventional cell and higher organism viability testing techniques. The feasibility of producing small instruments for toxicity detection and screening based on cellular dielectric responses is discussed.     

A density functional study of chemical,magnetic and thermodynamic properties of small palladium clusters

Structural, chemical, magnetic and thermodynamic properties of palladium clusters Pd_n with n = 2–11 are studied using density functional methods. The average bond length, entropy, enthalpy and polarisability are observed to increase as the cluster grows in size. The binding energy per atom also increases with cluster size. Stability function and atom addition energy change predict that Pd₄, Pd₆ and Pd₉ are relatively more stable than their neighbouring clusters. Electron affinity, electronegativity and electrophilicity values suggest that larger clusters have stronger tendency to accept electrons, thereby supporting the relative stability of Pd₄ and Pd₆. Chemical hardness is also seen to decrease with cluster size, which suggests that large clusters are more prone to changes in their electronic structure. The magnetic properties of these clusters are reported.     

Influence of training in the dark or light phase on physiologic and metabolic parameters of Wistar rats submitted to aerobic exercise

Light and dark phase training may influences rodents’ physiologic parameters because these animals have nocturnal habits. Thus, we verify the effects of the training in different photoperiods on metabolism and corporal composition of rats. Eighteen rats were divided into three groups – G1: non-trained; G2: trained in the light phase; G3: trained in the dark phase. Rats were allowed to swim for 60 min, five times per week during six weeks. Trained animals presented a smaller weight gain and fat percentage in carcass. Rats of G3 increased gastrocnemius relative weight. The adipocyte diameter of G3 rats was smaller than the other groups. The levels of the total cholesterol, low-density proteins, and triacylglycerols were decreased in animals of G2 while the glycemia was increased. Training in light phase provided more alterations in the blood biochemical profile while the training in the dark increased the gastrocnemius weight and decreased the diameter of the adipocyte.     

Biophysical studies of cholesterol effects on chromatin

Changes in chromatin structure regulate gene expression and genome maintenance. Molecules that bind to the nucleosome, the complex of DNA and histone proteins, are key modulators of chromatin structure. Previous work indicated that cholesterol, a ubiquitous cellular lipid, may bind to chromatin in vivo, suggesting a potential function for lipids in modulating chromatin architecture. However, the molecular mechanisms of cholesterol's action on chromatin structure have remained unclear. Here, we explored the biophysical impact of cholesterol on nucleosome and chromatin fibers reconstituted in vitro and characterized in silico the cholesterol binding to the nucleosome. Our findings support that cholesterol assists 10 and 30 nm chromatin formation and induces folding of long chromatin fibers as a result of direct interaction of the cholesterol to six nucleosomal binding sites.     

Effect of ubiquinol supplementation on biochemical and oxidative stress indexes after intense exercise in young athletes

Objectives: Physical exercise significantly impacts the biochemistry of the organism. Ubiquinone is a key component of the mitochondrial respiratory chain and ubiquinol, its reduced and active form, is an emerging molecule in sport nutrition. The aim of this study was to evaluate the effect of ubiquinol supplementation on biochemical and oxidative stress indexes after an intense bout of exercise.

Methods: 21 male young athletes (26?+?5 years of age) were randomized in two groups according to a double blind cross-over study, either supplemented with ubiquinol (200?mg/day) or placebo for 1 month. Blood was withdrawn before and after a single bout of intense exercise (40 min run at 85% maxHR). Physical performance, hematochemical parameters, ubiquinone/ubiquinol plasma content, intracellular reactive oxygen species (ROS) level, mitochondrial membrane depolarization, paraoxonase activity and oxidative DNA damage were analyzed.

Results: A single bout of intense exercise produced a significant increase in most hematochemical indexes, in particular CK and Mb while, on the contrary, normalized coenzyme Q₁₀ plasma content decreased significantly in all subjects. Ubiquinol supplementation prevented exercise-induced CoQ deprivation and decrease in paraoxonase activity. Moreover at a cellular level, in peripheral blood mononuclear cells, ubiquinol supplementation was associated with a significant decrease in cytosolic ROS while mitochondrial membrane potential and oxidative DNA damage remained unchanged.

Discussion: Data highlights a very rapid dynamic of CoQ depletion following intense exercise underlying an increased demand by the organism. Ubiquinol supplementation minimized exercise-induced depletion and enhanced plasma and cellular antioxidant levels but it was not able to improve physical performance indexes or markers of muscular damage.     

Structural analysis and antioxidative properties of mutan (water-insoluble glucan) and carboxymethyl mutan from Streptococcus mutans

Streptococcus mutans (MTCC 497) was grown anaerobically in acidic Brain heart infusion (BHI) medium with 15 % sucrose to produce cell-bound and extracellular water-insoluble polysaccharide mutan. Fourier transformed infrared (FTIR) and ¹³C NMR studies revealed a mixed linkage of α-1−3 and α-1–6 mutan with a production yield of 1.8 g/L. Mutan has a branched structure with a molecular weight (M_w) of 5654 Da. Water-insoluble mutan was carboxymethylated at 0.93 degrees of substitution. FTIR spectra with characteristic peaks at 1603 cm⁻¹ and 1418 cm⁻¹ due to symmetric and asymmetric vibrations of the COO- group confirmed carboxymethylation. Thermal gravimetric analysis showed that native mutan and carboxymethyl mutan exhibited higher thermal stability. Carboxymethylation enhanced solubility and antioxidative radical-scavenging activity. The in-vitro antioxidative radical scavenging analysis revealed 52 % and 47 % inhibition of DPPH and ABTS radicals.     

Odontocete bycatch and depredation in longline fisheries: A review of available literature and of potential solutions

Operational interactions between odontocetes (i.e., toothed whales) and longline gear are a global phenomenon that may threaten the conservation of odontocete populations and the economic viability of longline fisheries. This review attempts to define the issue, summarize the trends and geographical extent of its occurrence over the last half century, explore the potential impact on odontocetes and on fisheries, and describe potential acoustic and physical mitigation solutions. Reports of odontocete bycatch rates are highly variable (between 0.002 and 0.231 individuals killed per set) and at least 20 species may be involved. Information about marine mammal population size, migration patterns and life history characteristics are scarce, although at least one population may be in decline due to losses attributable to longline bycatch. Information about the financial impact of depredation on pelagic longline fisheries is also scarce, although estimates of daily fleet‐wide losses range between US$1,034 and US$8,495 (overall fleet income was not reported). Such biological and financial losses may be unsustainable. Recent developments in acoustic and physical mitigation technologies have yielded mixed results. Acoustic mitigation technologies have no moving parts, although require complex electronics. To date, they are insufficiently developed and their efficacy has been difficult to assess. Physical mitigation technologies generally require complex moving parts, although they are relatively simple to develop and assess. Further development and testing remains necessary before widespread implementation would be possible. Development of these approaches should be prioritized and a “toolbox” of various strategies and solutions should be compiled, because a single panacea to the problem is unlikely to emerge.     

Predictors of Weight Change in Middle‐aged and Old Men

Objective: Studies on weight change and mortality have yielded inconclusive results. This 10‐year prospective study was undertaken to improve understanding of factors affecting weight change. Research Methods and Procedures: The subjects were 1143 men, aged 36 to 88 years (mean, 53.3 years) at entry. A questionnaire was filled in at entry and at the end of the follow‐up with queries on weight, height, weight at the age of 20, physician‐diagnosed diseases, smoking, alcohol use, dietary habits, leisure physical activity, occupation, present occupational activity, living conditions (living alone or cohabiting), and former athletic status. Further information on morbidity was obtained from selected national registers. Factors predicting weight change during the study were identified by stepwise linear multiple regression analysis. Results: The mean 10‐year weight change was 0.8 (range, ?29 to +24) kg. Age at entry (β‐coefficient, ?0.17, SE 0.02), weight at entry (β, ?0.03, SE 0.01), diabetes at entry (β, ?3.55, SE 1.02), diabetes diagnosed after entry (β, ?3.94, SE 0.96), malignant cancer (β, ?1.60, SE 0.70), being a smoker (β, ?1.59, SE 0.48), and increased physical activity (β, ?1.27, SE 0.54) were significantly (p < 0.05) associated with weight loss in the final model. The model explained 13% of the variance of weight change. Discussion: The results emphasize the complexity of weight change. Some factors associated with weight change are apparently negatively, and some positively, associated with health. This could explain the equivocal findings on weight change and mortality in the literature.