Structure and properties of enzyme graft copolymers: Temperature effects on HRP immobilization mechanism

The possibility of obtaining immobilized horseradish peroxidase (HRP) materials with K'(m) values close to that of the native enzyme, but with good thermal stability, was investigated. The photochemical reaction was used as the immobilization methodology. Temperature and catalyst concentration were found to be the main parameters able to control the immobilization reaction mechanism more than type of functional monomer, polymer-matrix, and enzyme-polymer ratios. By carrying out the immobilization reaction at 35 degrees C and using either bisacryloylpiperazine (BAP) or hexhydro-1,3,5-triacryloyl-s-triazine (HTsT) as the functional monomer, materials with a good thermal stabilization (the retained activity after 240 min at 60 degrees C was between 65-25%) as well as kinetic constants (0.6-0.8 x 10(-4)M) similar to that of the free enzyme (0.57 x 10(-4)M) were obtained. Since low K'(m) values were obtained also using a high polymer content (pBAP copolymers, 25%; pHTsT copolymers, 30%) and neither limitation to substrate diffusion nor a reduction of the enzyme mobility was found, the enzyme should be linked to the matrix during the last steps of monomer polymerization, and it should have an external disposition with respect to the support.     

A phylogenomic resolution for the taxonomy of Aegean green lizards

Lacerta pamphylica and Lacerta trilineata are two currently recognized green lizard species with a historically problematic taxonomy. In cases of tangled phylogenies, next-generation sequencing and double-digest restriction-site-associated DNA protocols can provide a wealth of genomic data and resolve difficult taxonomic issues. Here, we generated genome-wide SNPs and mitochondrial sequences, and applied molecular species delimitation approaches to provide a stable taxonomy for the Aegean green lizards. Mitochondrial gene trees, genetic cluster delimitation and population structure analyses converged into recognizing the populations of (a) L. pamphylica, (b) east Aegean islands, Anatolia and Thrace (diplochondrodes lineage), (c) central Aegean islands (citrovittata), and (d) remaining Balkan populations and islands (trilineata), as separate clusters. Phylogenomic analyses revealed a split into two major clades, east and west of the Aegean Barrier, unambiguously showing a sister–clade relationship between pamphylica and diplochondrodes, rendering L. trilineata paraphyletic. Species delimitation models were tested in a Bayesian framework using the genomic SNPs: lumping all populations into a single ‘species’ had the lowest likelihood but the current taxonomy was also outperformed by all other models. All lines of evidence support the Pamphylian green lizard as a valid species; thus, east Aegean L. trilineata should also be considered a distinct species under the name Lacerta diplochondrodes. Finally, evidence from the mitochondrial and nuclear genomes is overwhelmingly in favour of recognizing the morphologically distinct Cycladian green lizards as a distinct species. We propose their elevation to full species under the name Lacerta citrovittata. All remaining insular and continental populations of the Balkan Peninsula represent the species L. trilineata.     

Mammary analogue secretory carcinoma of salivary gland diagnosed on submandibular gland cytology: A case report and review of the literature

Mammary analogue secretory carcinoma of the salivary glands has morphological shares molecular similarities to secretory carcinoma of the breast. Here, we report a 46‐year‐old woman who presented with a right submandibular gland mass. Fine needle aspiration differential diagnosis included oncocytosis, oncocytoma, acinic cell carcinoma and mammary analogue secretory carcinoma. We also review the current literature regarding clinical presentation and diagnostic workup of this entity.     

Semisynthetic Triterpenes Derived from Euphorbia officinarum as Plant Growth Promoters and Inducers of Disease Resistance

This study evaluated the effect of application of the semisynthetic triterpenes 3β-acetoxy-norlup-20-one (F4) and 3-chloro-4α,14α-dimethyl-5α-cholest-8-ene (F6) triterpene derivatives from Euphorbia officinarum on the growth of tomato seedlings under normal conditions and when challenged with the pathogens Verticillium dahliae and Agrobacterium tumefaciens. Foliar spray of F4 and F6 significantly improved growth rate, fresh weight, dry weight, and leaf area. In addition, they enhanced several physiological parameters including photosynthetic pigments, proline content, and nitrate reductase activity. Moreover, they induced H₂O₂ accumulation and increased the activity of several antioxidant enzymes such as catalase, ascorbate peroxidase, and guaiacol peroxidase. They also enhanced disease resistance against V. dahliae and A. tumefaciens. These results suggest that the two semisynthetic triterpenes represent new plant growth regulators and inducers of plant disease resistance.

     

Effect of organic and mineral fertilizers on N-use by wheat under different irrigation frequencies

A field trial was established in Errachidia, southern Morocco, to investigate the interaction between wheat residue management and mineral 15N-labelled ammonium sulphate, under different irrigation treatments, applied to wheat (Triticum durum var. Karim). In treatments I1, I2, I3 and I4, plots were irrigated every 10, 15, 21 and 30 days. Each plot contained three sub-plots that received three fertilization treatments: T1 received 42 kg N ha-1 of ammonium sulphate before seedling, 42 kg N ha-1 of ammonium sulphate labelled with 9.764 at % 15N excess at tillering and 84 N kg ha-1 of ammonium sulphate at flowering; T2 received 42 kg N ha-1 of ammonium sulphate labelled with 9.764 at % 15N excess at seedling, 42 kg N ha-1 at tillering and 42 kg N ha-1 at flowering; T3 received 4800 kg ha-1 of wheat residue labelled with 1.504 at % 15N excess and 42 kg N ha-1 of ammonium sulphate before seedling and 42 kg N ha-1 of ammonium sulphate at flowering. Nitrogen fertilization with 168 kg N ha-1 did no significantly increase grain and straw yields in comparison to the 126 kg N ha-1 application. The combination of the organic input and supplementary application of mineral fertilizer N has been found as a more attractive management option. For all irrigation treatments, the % recovery of N in the whole plant was higher in plants that received 15N at tillering (63%, 49% respectively for irrigation intervals between 10 and 30 d) than in plants that received 15N just after seeding (28% for irrigation each 10- and 30-d intervals). For the irrigation treatment each 10 and 15 days, the 15N was mainly recovered by the grain for all fertilization treatments, whereas for irrigation treatment each 30 days, the grain and straw recovered nearly equal amounts of fertilizer. For grain and straw of wheat, nitrogen in the plant derived from the fertilizer was low, while most of the N was derived from the soil for all irrigation and fertilization treatments. The % nitrogen in the plant derived from the fertilizer values showed no significant difference between the different plant parts. The results suggested a dominant influence of moisture availability on the fertilizer N uptake by wheat. Under dry conditions the losses of N can be allotted to denitrification and volatilisation.     

Diurnal and seasonal trends of water relations in five co-occurring chasmophytic species

The present study examines the seasonal and diurnal patterns of water management by plant species inhabiting the wall fissures of the ancient castle of Patras. Their water status (water potential), stomatal behaviour (leaf resistance and transpiration rate) as well as tissue water relations (turgor loss point, osmoregulation capability and cell wall elasticity) were recorded in relation to season, daytime and respective environmental conditions. Despite some minor deviations, all five species exhibited a water spending strategy with high diurnal transpiration rates through seasons, limited only by the generally low light intensities prevailing at the northwest-facing vertical walls. Progressive shortage of water during summer resulted in the reduction of transpiration in four species. Diurnal water losses caused a reduction of water potential until midday or dusk, which further decreased with the progress of the dry period. However, predawn water potential remained high through all seasons. The above finding, together with the high transpiration, was unexpected for species growing in a very hostile environment as far as water supply is regarded. It could be partly explained by the recently found ability of all five species to absorb dew from leaf surfaces. However, such plants should also possess mechanisms to take up water efficiently from a rather dry substrate. Indeed, pressure–volume analysis revealed substantial seasonal changes in osmoregulatory capacity and minor changes in cell wall elasticity of leaf tissue. Both changes facilitate (the mechanism differs) water uptake from the wall-fissure substrate during the dry period. Although the relative contribution of the two mechanisms was different, they both allowed plants to maintain turgor and thus growth throughout their growing season.     

Setup and use of a two-laser multiphoton microscope for multichannel intravital fluorescence imaging

Characterizing biological mechanisms dependent upon the interaction of many cell types in vivo requires both multiphoton microscope systems capable of expanding the number and types of fluorophores that can be imaged simultaneously while removing the wavelength and tunability restrictions of existing systems, and enhanced software for extracting critical cellular parameters from voluminous 4D data sets. We present a procedure for constructing a two-laser multiphoton microscope that extends the wavelength range of excitation light, expands the number of simultaneously usable fluorophores and markedly increases signal to noise via 'over-clocking' of detection. We also utilize a custom-written software plug-in that simplifies the quantitative tracking and analysis of 4D intravital image data. We begin by describing the optics, hardware, electronics and software required, and finally the use of the plug-in for analysis. We demonstrate the use of the setup and plug-in by presenting data collected via intravital imaging of a mouse model of breast cancer. The procedure may be completed in ～24 h.