Selected Contribution: Pulmonary hypertension in mice following intermittent hypoxia.

Sleep apnea (intermittent periods of hypoxia with or without hypercapnia) is associated with systemic hypertension and increased mortality from cardiovascular disease, but the relationship to pulmonary hypertension is uncertain. Previous studies on intermittent hypoxia (IH) in rats that demonstrated pulmonary hypertension utilized relatively long periods of hypoxia. Recent studies that utilized brief periods of hypoxia have conflicting reports of right ventricular (RV) hypertrophy. In addition, many studies have not measured pulmonary hemodynamics to asses the severity of pulmonary hypertension in vivo. Given the increasing availability of genetically engineered mice and the need to establish a rodent model of IH-induced pulmonary hypertension, we studied the effect of IH (2-min cycles of 10% and 21% O2, 8 h/day, 4 wk) on wild-type mice, correlating in vivo measurements of pulmonary hypertension with RV mass and pulmonary vascular remodeling. RV systolic pressure was increased after IH (36 +/- 0.9 mmHg) compared with normoxia (29.5 +/- 0.6) but was lower than continuous hypoxia (44.2 +/- 3.4). RV mass [RV-to-(left ventricle plus septum) ratio] correlated with pressure measurements (IH = 0.27 +/- 0.02, normoxia = 0.22 +/- 0.01, and continuous hypoxia = 0.34 +/- 0.01). Hematocrits were also elevated after IH and continuous hypoxia (56 +/- 1.6 and 54 +/- 1.1 vs. 44.3 +/- 0.5%). Evidence of neomuscularization of the distal pulmonary circulation was found after IH and continuous hypoxia. We conclude that mice develop pulmonary hypertension following IH, representing a possible animal model of pulmonary hypertension in response to the repetitive hypoxia-reoxygenation of sleep apnea.     

What paths do advantageous alleles take during short‐term evolutionary change?

Combining experimental evolution with whole‐genome resequencing is a promising new strategy for investigating the dynamics of evolutionary change. Published studies that have resequenced laboratory‐selected populations of sexual organisms have typically focused on populations sampled at the end of an evolution experiment. These studies have attempted to associate particular alleles with phenotypic change and attempted to distinguish between different theoretical models of adaptation. However, neither the population used to initiate the experiment nor multiple time points sampled during the evolutionary trajectory are generally available for examination. In this issue of Molecular Ecology, Orozco‐terWengel et al. (2012) take a significant step forward by estimating genome‐wide allele frequencies at the start, 15 generations into and at the end of a 37‐generation Drosophila experimental evolution study. The authors identify regions of the genome that have responded to laboratory selection and describe the temporal dynamics of allele frequency change. They identify two common trajectories for putatively adaptive alleles: alleles either gradually increase in frequency throughout the entire 37 generations or alleles plateau at a new frequency by generation 15. The identification of complex trajectories of alleles under selection contributes to a growing body of literature suggesting that simple models of adaptation, whereby beneficial alleles arise and increase in frequency unimpeded until they become fixed, may not adequately describe short‐term response to selection.     

Clinical mass spectrometry in neuroscience. Proteomics and peptidomics.

In this review we discuss the merits and drawbacks with the use of proteomic and peptidomic strategies for identification of proteins and peptides in their multidimensional interactions in complex biological processes. The progress in proteomics and peptidomics during the last years offer us new challenges to study changes in the protein and peptide synthesis. These strategies also offer new tools to follow post-translational modifications and other disturbed chemical processes that may be indicative of pathophysiological alteration(s). Furthermore these techniques can contribute to improvements in the diagnosis and therapy of neurodegenerative diseases, such as Alzheimer's disease, and psychiatric diseases, as depression and post traumatic stress disorders. We also consider different practical aspects of the applications of mass spectrometry in clinical neuroscience, illustrated by example from our laboratories. The new proteomic and peptidomic strategies will further enable the progress for clinical neuroscience research.     

Plasmodesmata signaling: many roles, sophisticated statutes.

Recent advances have increased our understanding of plasmodesmata function, their architecture as it relates to signaling capacity, the temporal and spatial regulation of their permeability, and their roles in systemic transport of macromolecules, non-cell autonomous development, and, potentially, plant defense.     

Preface

Molecular and Cellular Biochemistry -     

Biodegradation of BTEX by bacteria on powdered activated carbon

Aerobic degradation of a mixture of benzene, toluene, ethylbenzene and the mixed xylenes (BTEX) by a mixed bacterial population was studied in a continuously fed, completely mixed bioreactor in the presence of powdered activated carbon (PAC). Adsorption was characterized in the presence and in the absence of bacteria on PAC, and the affinity of virgin PAC to individual BTEX components was shown to be inversely correlated to their solubility in water. Bacteria colonizing the PAC particles are essential for simultaneous adsorption–biodegradation processes. In order to restrict biofilm formation and thereby mass transfer resistance of pollutants from the bulk to the PAC, the slurry was recirculated in the reactor system using a high shear pump. The bacteria on the PAC surface were constantly in a flux between the adsorbed and free phase, a phenomenon that prevented the formation of a biofilm on the PAC surface and thereby extended the life of the PAC.     

Non-linear control of continuous bioreactors

Control of bioreactors has achieved importance in the recent years. This may be due to the fact that they are difficult to control which may be attributed to its nonlinear dynamic behavior. The model parameters of the bioreactor also vary in an unpredictable manner. The complexity of the biochemical processes inhibits the accurate modeling and also the lack of suitable sensors make the process state difficult to characterize. Considerable emphasis has been placed on the control of fed-batch fermentors because of their prevalence in industries. However, when production of biomass is to be optimized, continuous operation is desirable. Several procedures are available for the nonlinear control of processes, viz., differential geometric approach, internal model control approach, reference synthesis technique, predictive control design, etc., but the major disadvantage of these approaches is the computational time required to perform the prediction optimization. In this study, a nonlinear controller based on a polynomial discrete time model (NARMAX) is evaluated for its performance on a fermentor. It can be shown that a nonlinear self-tuning controller based on NARMAX model can be extended to the control of fermentors. The response is smooth for both load and setpoint changes even when process parameters are assumed to be zero and uncertainty in parameters are present and in the presence of controller constraints. The control action can be made more or less robust by changing the design parameters appropriately. Therefore, nonlinear self-tuning controller is suitable for control of industrial processes.     

The catalytic activities of DNA topoisomerase II are most closely associated with the DNA cleavage/religation steps.

DNA topoisomerase II regulates the three-dimensional organisation of DNA and is the principal target of many important anticancer and antimicrobial agents. These drugs usually act on the DNA cleavage/religation steps of the catalytic cycle resulting in accumulation of covalent DNA-topoisomerase II complexes. We have studied the different steps of the catalytic cycle as a function of salt concentration, which is a classical way to evaluate the biochemical properties of proteins. The results show that the catalytic activity of topoisomerase II follows a bell-shaped curve with optimum between 100 and 225 mM KCl. No straight-forward correlation exists between DNA binding and catalytic activity. The highest levels of drug-induced covalent DNA-topoisomerase II complexes are observed between 100 and 150 mM KCl. Remarkably, at salt concentrations between 150 mM and 225 mM KCl, topoisomerase II is converted into a drug-resistant form with greatly reduced levels of drug-induced DNA-topoisomerase II complexes. This is due to efficient religation rather than to absence of DNA cleavage as witnessed by relaxation of the supercoiled DNA substrate. In the absence of DNA, ATP hydrolysis is strongest at low salt concentrations. Unexpectedly, the addition of DNA stimulates ATP hydrolysis at 100 and 150 mM KCl, but has little or no effect below 100 mM KCl in spite of strong non-covalent DNA binding at these salt concentrations. Therefore, DNA-stimulated ATP hydrolysis appears to be associated with covalent rather than non-covalent binding of DNA to topoisomerase II. Taken together, the results suggest that it is the DNA cleavage/religation steps that are most closely associated with the catalytic activities of topoisomerase II providing a unifying theme for the biological and pharmacological modulation of this enzyme.