A spin label study of erythrocyte membranes during simulation of freezing

Summary Human erythrocytes were labeled with stearic acid spin labels, and no change was detected in membrane fluidity under hyperosmotic stress, going from isotonicity to about 3000 mOsm. Intact erythrocytes labeled with an androstane spin label and submitted to simulation of freezing show the onset of irreversible structural breakdown occurring in a saline solution at 2,000 mOsm. Ghosts labeled with maleimide spin label (4-maleimide-2,2,6,6-tetramethylpiperidinooxyl) when submitted to solutions of increasing osmolalities (pH 7.4), exhibit protein conformational changes that are irreversible after a simulated freeze-thaw cycle. After sonication of maleimide spin-labeled ghosts, membrane buried sulfhydryl groups become exposed. Such preparations showed behavior similar to the unsonicated when in saline hyperosmolal medium (pH 7.4). Such results suggest the ionic strength of the medium as the determining factor of the detected conformational changes. Maleimide spin-labeled ghosts in 300 mOsm saline solution (pH 7.4) were treated with ascorbic acid (spin destruction of nitroxides), and the kinetic analysis indicates that 65% of the labeled sites are located at the external interface of the membrane or in hydrophilic channels. Deformation and rearrangements of membrane components in solutions of increasing osmolalities apparently are related to protein conformational changes, on the outside surface of erythrocyte membranes, with a significant amount being structurally dissociated of lipids.     

Visual Evoked Cortical Potential (VECP) Elicited by Sinusoidal Gratings Controlled by Pseudo-Random Stimulation

The contributions of contrast detection mechanisms to the visual cortical evoked potential (VECP) have been investigated studying the contrast-response and spatial frequency-response functions. Previously, the use of m-sequences for stimulus control has been almost restricted to multifocal electrophysiology stimulation and, in some aspects, it substantially differs from conventional VECPs. Single stimulation with spatial contrast temporally controlled by m-sequences has not been extensively tested or compared to multifocal techniques. Our purpose was to evaluate the influence of spatial frequency and contrast of sinusoidal gratings on the VECP elicited by pseudo-random stimulation. Nine normal subjects were stimulated by achromatic sinusoidal gratings driven by pseudo random binary m-sequence at seven spatial frequencies (0.4–10 cpd) and three stimulus sizes (4°, 8°, and 16° of visual angle). At 8° subtence, six contrast levels were used (3.12–99%). The first order kernel (K1) did not provide a consistent measurable signal across spatial frequencies and contrasts that were tested–signal was very small or absent–while the second order kernel first (K2.1) and second (K2.2) slices exhibited reliable responses for the stimulus range. The main differences between results obtained with the K2.1 and K2.2 were in the contrast gain as measured in the amplitude versus contrast and amplitude versus spatial frequency functions. The results indicated that K2.1 was dominated by M-pathway, but for some stimulus condition some P-pathway contribution could be found, while the second slice reflected the P-pathway contribution. The present work extended previous findings of the visual pathways contribution to VECP elicited by pseudorandom stimulation for a wider range of spatial frequencies.     

External coincidence and photoperiodic time measurement in the spider mite Tetranychus urticae

The incidence of diapause in the spider mite Tetranychus urticae was predicted for various photoperiodic regimes, according to the external coincidence model of photoperiodic time measurement. A phase response curve was constructed for the hypothetical photoperiodic oscillator in these mites: entrainment of this photoperiodic oscillator to a variety of ‘complete’ and ‘skeleton’ photoperiods was calculated using a transformation method for circadian rhythms. The external coincidence model proved adequate to describe experimental results with T. urticae in ‘complete’ photoperiods (T = 24 hr), symmetrical ‘skeleton’ photoperiods (T = 24 hr), asymmetrical ‘skeleton’ photoperiods (T = 24 hr) (night-interruption experiments), and ‘resonance’ experiments, in which the light component of a light/dark cycle was held constant at 8 hr and the dark component was varied over a wide range in successive experiments, providing cycles with period lengths up to 92 hr. The external coincidence model proved inadequate to explain results obtained in a ‘T-experiment’ with T. urticae comprising 1 hr pulses of light in a cycle of LD1:17.5 (T = 18.5 hr) with the first pulse of the train starting at different circadian phases. The validity and limitations of the external coincidence model as an explanation of photoperiodic time measurement in T. urticae are discussed in view of the above results.     

Beyond the green: assessing quarry restoration success through plant and beetle communities

In assessing the effectiveness of ecological restoration actions, outcomes evaluation using a multi‐taxa approach can greatly contribute to a clearer understanding of their success/failure. Since comprehensive biodiversity assessments are rarely possible, choosing taxa groups that are indicative of the ecosystem's structural and functional recovery is of major importance. Our goal was to evaluate the success of revegetation actions performed in a Mediterranean limestone quarry, using plants and epigean beetles as indicators. We compared their abundance, diversity, and community composition between revegetated sites aged 5, 13, and 19 years and a natural reference. Total plant cover significantly increased with restoration age and quickly reached reference values. However, native woody species cover dropped in the oldest site, while non‐native species became dominant. The abundance of beetles was always lower in restoration sites when compared to the reference, increasing with age, although not significantly. The richness of both plant species and beetle families was lower in restoration sites and did not show any trend towards the reference values. Finally, using nonmetric multidimensional scaling, the composition of plant and beetle communities from restoration sites showed a clear separation from the reference. Restoration efforts have successfully modified post‐quarry sites, but considerable differences remain, probably largely related to the use of the non‐native species Pinus halepensis in restoration plans. P. halepensis high cover in restoration sites greatly affects the structure of the ecosystem, and most likely its functioning too, as well as related ecosystem services, causing divergence from the reference values and compromising restoration success.     

Passive restoration in Araucaria Forest: useful ecological indicators in monitoring successional advancement in exotic tree plantation landscapes

Monitoring successional advancement is a complex field involving a constant search for applied ecological indicators which facilitate monitoring of secondary forests for both active and passive restoration. In this study, the authors investigate the successional advancement of floristics and tree structure within Araucaria Forest (AF) fragments under passive restoration in a context where exotic tree plantations (mainly Pinus L. genus) dominate the landscape. The ecological indicators used were floristic dissimilarity (β‐diversity inferences), indicator species, ecological groups of species, basal area, and species abundance distribution (SAD) models (α‐diversity inferences). A total of 182 tree species belonging to 91 genera and 43 botanical families were identified. A high β diversity was verified for which each site has indicator species (for the locations CD—Dicksonia sellowiana; CO—Cryptocarya aschersoniana; and PG—Pinus taeda), where pioneer species contributed to much of the abundance. Different SAD models are useful for describing passive restoration sites in exotic tree plantation landscapes, namely Lognormal, Mandelbrot, and Zipf. SAD models together with basal area, taxonomic group (e.g. Myrtaceae assemblage), and tree abundance in ecological groups are strategic ecological indicators for monitoring successional advancement in AF.     

Bioreactor production of 2,3-butanediol by Pantoea agglomerans using soybean hull acid hydrolysate as substrate

Production of 2,3-butanediol (2,3-BD) by Pantoea agglomerans strain BL1 was investigated using soybean hull hydrolysate as substrate in batch reactors. The cultivation media consisted of a mixture of xylose, arabinose, and glucose, obtained from the hemicellulosic fraction of the soybean hull biomass. We evaluated the influence of oxygen supply, pH control, and media supplementation on the growth kinetics of the microorganism and on 2,3-BD production. P. agglomerans BL1 was able to simultaneously metabolize all three monosaccharides present in the broth, with average conversions of 75% after 48 h of cultivation. The influence of aeration conditions employed demonstrated the mixed acid pathway of 2,3-BD formation by enterobacteria. Under fully aerated conditions (2 vvm of air), up to 14.02 g L⁻¹ of 2.3-BD in 12 h of cultivation were produced, corresponding to yields of 0.53 g g⁻¹ and a productivity of 1.17 g L⁻¹ h⁻¹, the best results achieved. These results suggest the production potential of 2,3-BD by P. agglomerans BL1, which has been recently isolated from an environmental consortium. The present work proposes a solution for the usage of the hemicellulosic fraction of agroindustry biomasses, carbohydrates whose utilization are not commonly addressed in bioprocess.