2‐D PAGE and MS analysis of proteins from formalin‐fixed,paraffin‐embedded tissues

In the past decade, encouraging results have been obtained in extraction and analysis of proteins from formalin‐fixed, paraffin‐embedded (FFPE) tissues. However, 2‐D PAGE protein maps with satisfactory proteomic information and comparability to fresh tissues have never been described to date. In the present study, we report 2‐D PAGE separation and MS identification of full‐length proteins extracted from FFPE skeletal muscle tissue. The 2‐D protein profiles obtained from FFPE tissues could be matched to those achieved from frozen tissues replicates. Up to 250 spots were clearly detected in 2‐D maps of proteins from FFPE tissue following standard mass‐compatible silver staining. Protein spots from both FFPE and frozen tissue 2‐D gels were excised, subjected to in situ hydrolysis, and identified by MS analysis. Matched spots produced matched protein identifications. Moreover, 2‐D protein maps from FFPE tissues were successfully subjected to Western immunoblotting, producing comparable results to fresh‐frozen tissues. In conclusion, this study provides evidence that, when adequately extracted, full‐length proteins from FFPE tissues might be suitable to 2‐D PAGE‐MS analysis, allowing differential proteomic studies on the vast existing archives of healthy and pathological‐fixed tissues.     

Productivity: key factor affecting grazing exclusion effects on vegetation and soil

In this study, we inquire into the effects of short-term goat grazing abandonment on plant species and functional composition, bare ground and net primary productivity (NPP) in two traditionally grazed pastures located in the Canarian Network of Natural Protected Areas and the Natura 2000 Network. In addition, we analyse soil chemical properties, biomass tannin content and energetic value to find out how grazing abandonment affects soil fertility and forage quality of these agroecosystems. Grazing exclusion effects on plant species and functional composition, as well as on soil fertility depended on the productivity of the studied pasture. Erect forbs and shrubs (endemic to Macaronesian region and native) were favoured by grazing removal in the most productive pasture, while soil fertility decreased in the driest and least productive site. An increase in NPP after exclusion was consistent among study sites. Although we consider goat grazing as necessary for maintaining traditional agroecosystems, we also suggest controlling it over time, allowing some periods of rest to give endemic shrub species time to recover from near propagule sources.     

Regulation of colony morphology and biofilm formation in Shewanella algae

Bacterial colony morphology can reflect different physiological stages such as virulence or biofilm formation. In this work we used transposon mutagenesis to identify genes that alter colony morphology and cause differential Congo Red (CR) and Brilliant Blue G (BBG) binding in Shewanella algae, a marine indigenous bacterium and occasional human pathogen. Microscopic analysis of colonies formed by the wild-type strain S. algae CECT 5071 and three transposon integration mutants representing the diversity of colony morphotypes showed production of biofilm extracellular polymeric substances (EPS) and distinctive morphological alterations. Electrophoretic and chemical analyses of extracted EPS showed differential patterns between strains, although the targets of CR and BBG binding remain to be identified. Galactose and galactosamine were the preponderant sugars in the colony biofilm EPS of S. algae. Surface-associated biofilm formation of transposon integration mutants was not directly correlated with a distinct colony morphotype. The hybrid sensor histidine kinase BarA abrogated surface-associated biofilm formation. Ectopic expression of the kinase and mutants in the phosphorelay cascade partially recovered biofilm formation. Altogether, this work provides the basic analysis to subsequently address the complex and intertwined networks regulating colony morphology and biofilm formation in this poorly understood species.     

Effect of temperature and ripening stages on membrane integrity of fresh-cut tomatoes

The shelf-life of fresh-cut tomatoes mainly depends on loss of tissue integrity and firmness that occurs also in intact fruits after long-term cold storage due to chilling injury. Round-fruit tomatoes (Solanum lycopersicum L.) cv. Jama were stored in 1.1-L plastic (polyethylene) fresh-cut produce containers as 10.0-mm-thick tomato slices and as intact tomatoes at 4 ± 0.5 °C. The aim of this work was to study the loss of membrane integrity and biochemical processes involved in membrane disruption. Electrolyte leakage and lipid peroxidation were studied at different stages of maturity: mature green, pink (PK), fully ripe and two different storage temperatures: 4 and 15 °C. The tomato slices of PK stage stored at 4 °C did not show changes for both parameters, while significant increase in membrane leakage and lipid peroxidation was observed at 15 °C, especially after 24 h of storage. The enzymes showed a simultaneous increase in their activities with a rise in electrolyte leakage and lipid peroxidation after 7 days of storage. Finally, phospholipase C (PLC) and phospholipase D (PLD) were investigated for intact fruit and tomato slices stored at 4 °C. The PLC had higher activity compared with PLD. In conclusion, the loss of membrane integrity in fresh-cut tomatoes is mainly affected by ripening stages, storage temperature and duration. The wounds enhance the PLC and PLD activities and they play a role late during storage.     

Autonomic Adaption to Clinical Simulation in Psychology Students: Teaching Applications

Simulation is used to facilitate new learning in a variety of situations. One application of simulation could be to help therapists gain therapeutic skills prior to seeing clients. This particular study was interested in measuring changes in stress response by looking at subjective and objective measures of distress (as measured by SUDS, HR, and HRV) over three sessions of simulated therapy. 16 second year psychology students participated in three sessions, and had their HR and HRV measured by Polar watches. Over the three sessions, there was a decrease in perceived distress, as measured by SUDS ratings. During and between sessions, there was inconclusive change in physiological parameters.     

Light response in seedlings of a temperate (<Emphasis Type="Italic">Quercus petraea</Emphasis>) and a sub-Mediterranean species (<Emphasis Type="Italic">Quercus pyrenaica</Emphasis>): contrasting ecological strategies as potential keys to regeneration performance in mixed marginal populations

In order to understand better the ecology of the temperate species Quercus petraea and the sub-Mediterranean species Quercus pyrenaica, two deciduous oaks, seedlings were raised in two contrasting light environments (SH, 5.3% full sunlight vs. HL, 70% full sunlight) for 2 years, and a subset of the SH seedlings were transferred to HL (SH–HL) in the summer of the second year. We predicted that Q. pyrenaica would behave more as a stress-tolerant species, with lower specific leaf area (SLA), allocation to leaf mass, and growth rate and less responsiveness to light in these metrics, than Q. petraea, presumed to be more competitive when resources, especially light and water, are abundant. Seedlings of Q. petraea had larger leaves with higher SLA, and exhibited a greater relative growth rate (RGR) in both SH and HL. They also displayed a higher proportion of biomass in stems (SMF), and a lower root to shoot ratio (R/S) in HL than those of Q. pyrenaica, which sprouted profusely, and had higher rates of photosynthesis (A_n) and stomatal conductance (g_wv), but lower whole-plant net assimilation rate (NAR). On exposure to a sudden increase in light, SH–HL seedlings of both species showed a short period of photoinhibition, but fully acclimated photosynthetic features within 46 days after transference; height, main stem diameter, RGR and NAR all increased at the end of the experiment compared to SH seedlings, with these increases more pronounced in Q. petraea. Observed differences in traits and responses to light confirmed a contrasting ecology at the seedling stage in Q. petraea and Q. pyrenaica in consonance with differences in their overall distribution. We discuss how the characteristics of Q. petraea may limit the availability of suitable regeneration niches to microsites of high-resource availability in marginal populations of Mediterranean climate, with potential negative consequences for its recruitment under predicted climatic changes.     

The structure of a galactan sulfate from the red seaweed Bostrychia montagnei

The sulfated, methylated galactan isolated from the red seaweed Bostrychia montagnei, showed an unusually narrow structural dispersion. This agaran has the defining linear backbone of alternating 3-linked beta-D-galactopyranosyl units and 4-linked alpha-L-galactopyranosyl and 3,6-anhydrogalactopyranosyl residues. The D-units have C-6 methylation, C-6 single stubs of xylopyranosyl and minor to trace amounts of (possible) C-6 linked single stubs of galactopyranosyl. These units are mainly sulfated on C-4 with lesser sulfation at C-6 and minor at C-2. The L-residues are mainly methylated on C-2 of the 3,6-anhydrogalactopyranosyl and sulfated on C-3 of the L-galactopyranosyl; minor amounts of 2,3- and 3,6-disulfated and 2-O-methyl or 2-O-glycosyl 3-sulfated L-galactopyranosyl were also found.     

An electromyographic analysis of two handwriting grasp patterns

BackgroundHandwriting is a fundamental skill needed for the development of daily-life activities during lifetime and can be performed using different forms to hold the writing object. In this study, we monitored the sEMG activity of trapezius, biceps brachii, extensor carpi radialis brevis and flexor digitorum superficialis during a handwriting task with two groups of subjects using different grasp patterns.Subjects and methodsTwenty-four university students (thirteen males and eleven females; mean age of 22.04 ± 2.8 years) were included in this study. We randomly invited 12 subjects that used the Dynamic Tripod grasp and 12 subjects that used the Static Tripod grasp.ResultsThe static tripod group showed statistically significant changes in the sEMG activity of trapezium and biceps brachii muscles during handwriting when compared to dynamic tripod group’s subjects. No significant differences were found in extensor carpi radialis brevis and flexor digitorum superficialis activities among the two groups.ConclusionThe findings in this study suggest an increased activity of proximal muscles among subjects using a transitional grasp, indicating potential higher energy expenditure and muscular harm with the maintenance of this motor pattern in handwriting tasks, especially during the progression in academic life.