The Newly Sequenced Genome of Pisum sativum Is Replete with Potential G-Quadruplex-Forming Sequences—Implications for Evolution and Biological Regulation

G-quadruplexes (G4s) have been long considered rare and physiologically unimportant in vitro curiosities, but recent methodological advances have proved their presence and functions in vivo. Moreover, in addition to their functional relevance in bacteria and animals, including humans, their importance has been recently demonstrated in evolutionarily distinct plant species. In this study, we analyzed the genome of Pisum sativum (garden pea, or the so-called green pea), a unique member of the Fabaceae family. Our results showed that this genome contained putative G4 sequences (PQSs). Interestingly, these PQSs were located nonrandomly in the nuclear genome. We also found PQSs in mitochondrial (mt) and chloroplast (cp) DNA, and we experimentally confirmed G4 formation for sequences found in these two organelles. The frequency of PQSs for nuclear DNA was 0.42 PQSs per thousand base pairs (kbp), in the same range as for cpDNA (0.53/kbp), but significantly lower than what was found for mitochondrial DNA (1.58/kbp). In the nuclear genome, PQSs were mainly associated with regulatory regions, including 5′UTRs, and upstream of the rRNA region. In contrast to genomic DNA, PQSs were located around RNA genes in cpDNA and mtDNA. Interestingly, PQSs were also associated with specific transposable elements such as TIR and LTR and around them, pointing to their role in their spreading in nuclear DNA. The nonrandom localization of PQSs uncovered their evolutionary and functional significance in the Pisum sativum genome.     

Empagliflozin Treatment Attenuates Hepatic Steatosis by Promoting White Adipose Expansion in Obese TallyHo Mice

Sodium-glucose co-transporters (SGLTs) serve to reabsorb glucose in the kidney. Recently, these transporters, mainly SGLT2, have emerged as new therapeutic targets for patients with diabetes and kidney disease; by inhibiting glucose reabsorption, they promote glycosuria, weight loss, and improve glucose tolerance. They have also been linked to cardiac protection and mitigation of liver injury. However, to date, the mechanism(s) by which SGLT2 inhibition promotes systemic improvements is not fully appreciated. Using an obese TallyHo mouse model which recapitulates the human condition of diabetes and nonalcoholic fatty liver disease (NAFLD), we sought to determine how modulation of renal glucose handling impacts liver structure and function. Apart from an attenuation of hyperglycemia, Empagliflozin was found to decrease circulating triglycerides and lipid accumulation in the liver in male TallyHo mice. This correlated with lowered hepatic cholesterol esters. Using in vivo MRI analysis, we further determined that the reduction in hepatic steatosis in male TallyHo mice was associated with an increase in nuchal white fat indicative of “healthy adipose expansion”. Notably, this whitening of the adipose came at the expense of brown adipose tissue. Collectively, these data indicate that the modulation of renal glucose handling has systemic effects and may be useful as a treatment option for NAFLD and steatohepatitis.     

Conception and construction of a Rasch-scaled measure for self-confidence in one's vocabulary ability

Although different theoretical approaches indicate the importance of self-confidence in one's ability in educational or counselling psychology, there are only a few standardized psychological instruments, these being mainly questionnaires, that focus on this construct. The idea pursued is to expand a traditional ability test and create an objective personality measure for self-confidence in one's ability. Empirical evidence will be provided as to whether it is possible to construct and cross validate this measure using the Rasch model. A two step procedure allowing for a self rating of one's knowledge of words was designed using 25 items from a general knowledge - vocabulary test. The results indicate that it is possible to construct and cross validate a measure for self-confidence in one's ability using the Rasch model. Additionally, it is shown that the measure for self-confidence in one's ability is independent from the general knowledge.     

Classification criteria and probability risk maps: limitations and perspectives

Delineation of polluted zones with respect to regulatory standards, accounting at the same time for the uncertainty of the estimated concentrations, relies on classification criteria that can lead to significantly different pollution risk maps, which, in turn, can depend on the regulatory standard itself. This paper reviews four popular classification criteria related to the violation of a probability threshold or a physical threshold, using annual (1996-2000) nitrogen dioxide concentrations from 40 air monitoring stations in Milan. The relative advantages and practical limitations of each criterion are discussed, and it is shown that some of the criteria are more appropriate for the problem at hand and that the choice of the criterion can be supported by the statistical distribution of the data and/or the regulatory standard. Finally, the polluted area is estimated over the different years and concentration thresholds using the appropriate risk maps as an additional source of uncertainty.     

Microfluidics: Nanostructuring of Titania Thin Films by a Combination of Microfluidics and Block‐Copolymer‐Based Sol–Gel Templating (Small 7/2011)

Sol–gel templating of titania thin films with the amphiphilic diblock copolymer poly(dimethyl siloxane)‐block‐methyl methacrylate poly(ethylene oxide) is combined with microfluidic technology to control the structure formation. Due to the laminar flow conditions in the microfluidic cell, a better control of the local composition of the reactive fluid is achieved. The resulting titania films exhibit mesopores and macropores, as determined with scanning electron microscopy, X‐ray reflectivity, and grazing incidence small angle X‐ray scattering. The titania morphology has three features that are beneficial for application in photovoltaics: 1) a large surface‐to‐volume ratio important for charge generation with disordered hexagonally arranged mesopores of 25 nm size and a film porosity of up to 0.79, 2) enhanced light scattering that enables the absorption of more light, and 3) a dense titania layer with a thickness of about 6 nm at the substrate (bottom electrode) to prevent short circuits. An optical characterization complements the structural investigation.     

Effect of internal curing by using superabsorbent polymers (SAP) on autogenous shrinkage and other properties of a high-performance fine-grained concrete: results of a RILEM round-robin test

The article presents the results of a round-robin test performed by 13 international research groups (representing fifteen institutions) in the framework of the activities of the RILEM Technical Committee 225-SAP “Applications of Superabsorbent Polymers in Concrete Construction”. Two commercially available SAP materials were used for internal curing of a high-performance, fine-grained concrete in combination with the addition of extra water. The concrete had the same mix composition in all laboratories involved but was composed of local materials. All found a considerable decrease in autogenous shrinkage attributable to internal curing. Also, with regard to the shrinkage-mitigating effect of both particular SAP materials, the results were consistent. This demonstrates that internal curing using SAP is a robust approach, working independently of some variations in the concretes’ raw materials, production process, or measuring technique. Furthermore, the effects of internal curing on other properties of concrete in its fresh and hardened states were investigated. These are consistent as well and expand considerably the existing data basis on properties of concrete materials containing SAP.     

Antimicrobial Peptides from the Aurein Family Form Ion‐Selective Pores in Bacillus subtilis

The mechanism of action of aurein 2.2 and aurein 2.3, antimicrobial peptides from the frog Litoria aurea, was investigated. Proteomic profiling of the Bacillus subtilis stress response indicates that the cell envelope is the main target for both aureins. Upon treatment, the cytoplasmic membrane depolarizes and cellular ATP levels decrease. Global element analysis shows that intracellular concentrations of certain metal ions (potassium, magnesium, iron, and manganese) strongly decrease. Selective translocation of some ions over others was demonstrated in vitro. The same set of ions also leaks from B. subtilis cells treated with sublethal concentrations of gramicidin S, MP196, and nisin. Aureins do not permeabilize the cell membrane for propidium iodide thus excluding formation of large, unspecific pores. Our data suggest that the aureins acts by forming small pores thereby causing membrane depolarization, and by triggering the release of certain metal ions thus disturbing cellular ion homeostasis.     

Detection of Proteinases in Saccharomyces cerevisiae by Flow Cytometry

The physiological state of a yeast population used for inoculation determines how rapidly the cells adapt to new environmental conditions, begin proliferating and utilising extract. The decision as to whether a yeast culture is suitable for re‐pitching should not be based only on viability determinations since this can be misleading. Increased proteolytic activity in a yeast population indicates the onset of senescence. A flow cytometric method has been developed for measuring a wide variety of proteinases in Saccharomyces cerevisiae employing a commercially available casein‐dye conjugate. The detection of intracellular proteinase activity gives an early indication of apoptotic events and allows improved assessment of the physiological state of a yeast population. This knowledge will assist the industry to optimize the selection of yeast and its subsequent fermentation performance. Yeast cell autolysis with all its negative consequences for beer quality and stability will thus be minimised.     

Phenolic compounds in tomatoes. Natural variations and effect of gamma-irradiation

A method for determination of p-hydroxybenzaldehyde, p-coumaric acid, ferulic acid, rutin and naringenin in tomatoes based on HPLC with coulometric electrode array detection was developed. After lyophilisation and extraction the compounds were separated isocratically (acetonitrile/buffer: 14:86, v:v, or acetonitrile/buffer: 27:73, v:v, flow rate: 0.8 ml/min) on reversed phase columns (LiChrospher 60 RP select B or Spherisorb ODS2) and detected at 16 respectively 10 working electrodes set at potentials between +50 and +750 mV against palladium reference electrodes. The natural content of these components showed a strong variation between different varieties, harvests and degrees of maturity. Gamma-irradiation reduced the concentration of the phenolic compounds markedly in all tomato samples investigated, however, this change was smaller than the naturally occurring differences. The irradiation products vanillin and eriodictyol, identified in standard solutions, could not be found in irradiated tomatoes.