The synergistic effect of carbon concentration and high temperature on lipid peroxidation in Peridinium gatunense

Lipid peroxidation in Peridinium samples taken from two differentdepths in Lake Kinneret fluctuated throughout the spring withan overall increasing trend. Samples from 0.5 and 5 m showeda similar peroxidation pattern, which was maximal after thefall off in algal biomass. The rapid decline in Peridinium biomasscoincided with ambient lake temperatures of 21–23C. Fattyacid composition profiles were similar at both depths, althoughafter the peak of the bloom, a significant increase in polyunsaturatedfatty acids and oleic acid was only found at 0.5 m, togetherwith a decrease in the percentage of polyunsaturated fatty acids.These effects were related to ambient light stress rather thana result of lipid peroxidation. Lake samples taken at differentperiods of the bloom and incubated at various temperatures showeddifferential peroxidation. Higher temperatures caused increasedlipid peroxidation, but this appeared to be dependent on thesampling period. Samples withdrawn from the lake at the beginningof the bloom showed little peroxidation after a 5 day incubationat 14C, room temperature (25C) or ambient lake temperature(16C) compared to mid-bloom samples in which there was a significantincrease in peroxidation when they were incubated at room temperature(25C) or ambient lake temperature (22C). Incubation at 14Cinhibited peroxidation; however, samples from mid-bloom againshowed enhanced peroxidation compared with those from the beginningof the bloom. These in situ results suggested a relationshipbetween temperature, another environmental variable during thebloom and lipid peroxidation in Peridinium. As total dissolvedinorganic carbon (DIC) concentrations fall significantly duringthe progress of the bloom and represent an important sourceof environmental stress, laboratory experiments were establishedto investigate the synergistic effect of temperature and carbonnutrition on lipid peroxidation in Peridinium cultures. Increasedtemperature alone caused a slight increase in lipid peroxidation,but this was greatly augmented by carbon limitation. Althoughcarbon limitation induced increased catalase activity, at highertemperatures activity declined after 48 h, allowing for thesubstantial increase in lipid peroxidation.     

Heat stress in the presence of low RNA polymerase activity increases chromosome copy number of Escherichia coli

Summary A temperature shift-up accompanied by a reduction in RNA polymerase activity in Escherichia coli causes an increased rate of initiation leading to a 1.7- to 2.2-fold increase in chromosome copy number. A temperature shift-up without a reduction in polymerase activity induces only a transient non-scheduled initiation of chromosome replication caused by heat shock with no detectable effect on chromosome copy number.     

Rendiconti Delle Sedute E Brevi Comunicazioni

Abstract

The development of effective strategies to rehabilitate compromised environments is one of the major challenges facing the postindustrial society, since most of the habitats are becoming progressively polluted due to indiscriminate discharge of contaminants generated by anthropogenic activities. Several aquatic photosynthetic organisms can be used to treat wastewaters in primary and tertiary treatments, including cyanobacteria, algae and higher plants. In this review, we summarize the results obtained in the remediation of polluted waters by photosynthetic organisms and discuss the future perspective of phytoremediation.     

Resolution of Two Sub-Populations of Conformers and Their Individual Dynamics by Time Resolved Ensemble Level FRET Measurements

Most active biopolymers are dynamic structures; thus, ensembles of such molecules should be characterized by distributions of intra- or intermolecular distances and their fast fluctuations. A method of choice to determine intramolecular distances is based on Förster resonance energy transfer (FRET) measurements. Major advances in such measurements were achieved by single molecule FRET measurements. Here, we show that by global analysis of the decay of the emission of both the donor and the acceptor it is also possible to resolve two sub-populations in a mixture of two ensembles of biopolymers by time resolved FRET (trFRET) measurements at the ensemble level. We show that two individual intramolecular distance distributions can be determined and characterized in terms of their individual means, full width at half maximum (FWHM), and two corresponding diffusion coefficients which reflect the rates of fast ns fluctuations within each sub-population. An important advantage of the ensemble level trFRET measurements is the ability to use low molecular weight small-sized probes and to determine nanosecond fluctuations of the distance between the probes. The limits of the possible resolution were first tested by simulation and then by preparation of mixtures of two model peptides. The first labeled polypeptide was a relatively rigid Pro₇ and the second polypeptide was a flexible molecule consisting of (Gly-Ser)₇ repeats. The end to end distance distributions and the diffusion coefficients of each peptide were determined. Global analysis of trFRET measurements of a series of mixtures of polypeptides recovered two end-to-end distance distributions and associated intramolecular diffusion coefficients, which were very close to those determined from each of the pure samples. This study is a proof of concept study demonstrating the power of ensemble level trFRET based methods in resolution of subpopulations in ensembles of flexible macromolecules.     

MzrA: a novel modulator of the EnvZ/OmpR two-component regulon

Analysis of suppressors that alleviate the acute envelope stress phenotype of a Δ bamB Δ degP strain of Escherichia coli identified a novel protein MzrA and pleiotropic envZ mutations. Genetic evidence shows that overexpression of MzrA – formerly known as YqjB and EcfM – modulates the activity of EnvZ/OmpR similarly to pleiotropic EnvZ mutants and alter porin expression. However, porin expression in strains devoid of MzrA or overexpressing it is still sensitive to medium osmolarity, pH and procaine, all of which modulate EnvZ/OmpR activities. Thus, MzrA appears to alter the output of the EnvZ/OmpR system but not its ability to receive and respond to various environmental signals. Localization and topology experiments indicate that MzrA is a type II membrane protein, with its N-terminus exposed in the cytoplasm and C-terminus in the periplasm. Bacterial two-hybrid experiments determined that MzrA specifically interacts with EnvZ but not with OmpR or the related membrane sensor kinase, CpxA. This and additional genetic and biochemical evidence suggest that the interaction of MzrA with EnvZ would either enhance EnvZ's kinase activity or reduce its phosphatase activity, thus elevating the steady state levels of OmpR∼P. Furthermore, our data show that MzrA links the two-component envelope stress response regulators, CpxA/CpxR and EnvZ/OmpR.     

Chronic hypoxic decreases in soluble guanylate cyclase protein and enzyme activity are age dependent in fetal and adult ovine carotid arteries.

The present study tests the hypothesis that chronic hypoxia enhances reactivity to nitric oxide (NO) through age-dependent increases in soluble guanylate cyclase (sGC) and protein kinase G (PKG) activity. In term fetal and adult ovine carotids, chronic hypoxia had no significant effect on mRNA levels for the beta1-subunit of sGC, but depressed sGC abundance by 16% in fetal and 50% in adult arteries, through possible depression of rates of mRNA translation (15% in fetal and 50% in adult) and/or increased protein turnover. Chronic hypoxia also depressed the catalytic activity of sGC, but only in fetal arteries (63%). Total sGC activity was reduced by chronic hypoxia in both fetal (69%) and adult (37%) carotid homogenates, but this effect was not observed in intact arteries when sGC activity was measured by timed accumulation of cGMP. In intact arteries treated with 300 microM 3-isobutyl-1-methylxanthine (IBMX), chronic hypoxia dramatically enhanced sGC activity in fetal (186%) but not adult (89%) arteries. This latter observation suggests that homogenization either removed an sGC activator, released an sGC inhibitor, or altered the phosphorylation state of the enzyme, resulting in reduced activity. In the absence of IBMX, chronic hypoxia had no significant effect on rates of cGMP accumulation. Chronic hypoxia also depressed the ability of the cGMP analog, 8-(p-chlorophenylthio)-cGMP, to promote vasorelaxation in both fetal (8%) and adult (12%) arteries. Together, these results emphasize the fact that intact and homogenized artery studies of sGC activity do not always yield equivalent results. The results further suggest that enhancement of reactivity to NO by chronic hypoxia must occur upstream of PKG and can only be possible if changes in cGMP occurred in functional compartments that afforded either temporal or chemical protection to the actions of phosphodiesterase. The range and age dependence of hypoxic effects observed also suggest that some responses to hypoxia must be compensatory and homeostatic, with reactivity to NO as the primary regulated variable.     

Cinnamaldehyde regulates H2O 2-induced skeletal muscle atrophy by ameliorating the proteolytic and antioxidant defense systems

Skeletal muscle atrophy/wasting is associated with impaired protein metabolism in diverse physiological and pathophysiological conditions. Elevated levels of reactive oxygen species (ROS), disturbed redox status, and weakened antioxidant defense system are the major contributing factors toward atrophy. Regulation of protein metabolism by controlling ROS levels and its associated catabolic pathways may help in treating atrophy and related clinical conditions. Although cinnamaldehyde (CNA) enjoys the established status of antioxidant and its role in ROS management is reported, impact of CNA on skeletal muscle atrophy and related pathways is still unexplored. In the current study, the impact of CNA on C2C12 myotubes and the possible protection of cultured cells from H ₂O ₂-induced atrophy is examined. Myotubes were treated with H ₂O ₂ in the presence and absence of CNA and the changes in the antioxidative, proteolytic systems, and mitochondrial functions were scored. Morphological analysis showed significant protective effects of CNA on length, diameter, and nuclei fusion index of myotubes. The evaluation of biochemical markers of atrophy; creatine kinase, lactate dehydrogenase, succinate dehydrogenase along with the study of muscle-specific structural protein (i.e., myosin heavy chain-fast [MHCf] type) showed significant protection of proteins by CNA. CNA pretreatment not only checked the activation of proteolytic systems (ubiquitin-proteasome E3-ligases [MuRF1/Atrogin1]), autophagy [Beclin1/LC3B], cathepsin L, calpain, caspase), but also prevented any alteration in the activities of antioxidative defense enzymes (catalase, glutathione- S-transferase, glutathione-peroxidase, superoxide dismutase, glutathione reductase). The results suggest that CNA protects myotubes from H ₂O ₂-induced atrophy by inhibiting/resisting the amendments in proteolytic systems and maintains cellular redox-balance.     

Characterization and Evaluation of 5-Fluorouracil-Loaded Solid Lipid Nanoparticles Prepared via a Temperature-Modulated Solidification Technique

The aim of this research was to advance solid lipid nanoparticle (SLN) preparation methodology by preparing glyceryl monostearate (GMS) nanoparticles using a temperature-modulated solidification process. The technique was reproducible and prepared nanoparticles without the need of organic solvents. An anticancer agent, 5-fluorouracil (5-FU), was incorporated in the SLNs. The SLNs were characterized by particle size analysis, zeta potential analysis, differential scanning calorimetry (DSC), infrared spectroscopy, atomic force microscopy (AFM), transmission electron microscopy (TEM), drug encapsulation efficiency, in vitro drug release, and in vitro cell viability studies. Particle size of the SLN dispersion was below 100 nm, and that of redispersed lyophilizates was ~500 nm. DSC and infrared spectroscopy suggested that the degree of crystallinity did not decrease appreciably when compared to GMS. TEM and AFM images showed well-defined spherical to oval particles. The drug encapsulation efficiency was found to be approximately 46%. In vitro drug release studies showed that 80% of the encapsulated drug was released within 1 h. In vitro cell cultures were biocompatible with blank SLNs but demonstrated concentration-dependent changes in cell viability to 5-FU-loaded SLNs. The 5-FU-loaded SLNs can potentially be utilized in an anticancer drug delivery system.KEY WORDS: atomic force microscopy, calorimetry (DSC), FTIR, particle size, solid lipid nanoparticles     

Dynamics of a Kalahari long-lived mega-lake system: hydromorphological and limnological changes in the Makgadikgadi Basin (Botswana) during the terminal 50 ka

The Kalahari features a long-lived lacustrine system which may exist since the Early Pleistocene. The emergence of an extant cichlid fish radiation from this (palaeo-) lake during the Middle Pleistocene indicates an ancient lake character. The early history of the system remains speculative, but it is established that lake extensions matching modern Lake Victoria in size have occurred during the Late Pleistocene. It has been assumed that the hydrographical dynamics chiefly depended on the inflow from the Okavango River and thus on ITCZ-controlled precipitation. Our studies, which focused the hydromorphological and palaeolimnological development of the Makgadikgadi Basin during the last 50 ka, suggest that from c. 46–16 ka it did not receive water from the Okavango River but from palaeo-rivers located in the northern and south-western catchment. A northward shift of the winter rainfall zone during the Last Glacial Maximum sustained a high lake level for a period of c. 6 ka. During Heinrich Event 1 (17–16 ka) the lake probably desiccated abruptly and completely. Higher lake levels, controlled by water from the Okavango river system, were reached again during the Holocene before the lake dried up in the middle of the last millennium.