Comparing sleep-loss sleepiness and sleep inertia: lapses make the difference

To compare the behavioral effects of sleep-loss sleepiness (performance impairment due to sleep loss) and sleep inertia (period of impaired performance that follows awakening), mean response latencies and number of lapses from a visual simple reaction-time task were analyzed. Three experimental conditions were designed to manipulate sleepiness and sleep-inertia levels: uninterrupted sleep, partial sleep reduction, and total sleep deprivation. Each condition included two consecutive nights (the first always a night of uninterrupted sleep, and the second either a night of uninterrupted sleep, a night when sleep was reduced to 3 h, or a night of total sleep deprivation), as well as two days in which performance was assessed at 10 different time points (08:00, 08:30, 09:00, 09:30, 10:00, 11:00, 14:00, 17:00, 20:00, and 23:00 h). From 08:00 to 09:00 h, reaction times in the partial sleep-reduction and total sleep-deprivation conditions were at a similar level and were slower than those observed in the uninterrupted sleep condition. In the same time period, the frequency of lapses in the total sleep-deprivation condition was higher than in the partial sleep-reduction condition, while this latter condition never differed from the uninterrupted sleep condition. The results indicate that both sleep inertia and sleep-loss sleepiness lead to an increase in response latencies, but only extreme sleepiness leads to an increase in lapse frequency. We conclude that while reaction times slow as a result of both sleep inertia and sleep-loss sleepiness, lapses appear to be a specific feature of sleep-loss sleepiness.     

Complexes of 2-acetyl-gamma-butyrolactone and 2-furancarbaldehyde thiosemicarbazones: antibacterial and antifungal activity

Cobalt, nickel, copper and zinc coordination compounds of two thiosemicarbazones with general composition ML₂ (L: monodeprotonated ligand corresponding to 2-acetyl-γ-butyrolactone thiosemicarbazone, HL¹, and 2-furancarbaldehyde thiosemicarbazone, HL²) and also complexes with general composition MCl₂(HL²) were synthesized (except [NiCl₂(HL²)] and [Co(L²)₂]). The interaction of CuCl₂ with HL² gave [CuCl(HL²)], a copper(I) complex. The ligands and metal complexes were characterized by IR, ¹H and ¹³C NMR spectroscopy, and magnetic susceptibility measurements. The crystal structure of [Ni(L²)₂] · 2dmso was determined and a trans-square planar coordination of the two κ²-N,S chelate rings forming polymeric strips through H-bonds with dmso was observed. Actually, in all the reported complexes both ligands behaved as κ²-N,S chelates, except in the case of [Co(L¹)₂] in which HL¹ is tridentate κ³-N,S,O. The antimicrobial properties of all compounds were studied using a wide spectrum of bacterial and fungal strains. The copper complexes of HL² were the most active against all strains, including dermatophytes and phytopathogenic fungi. Most of the studied compounds, especially [Cu(L¹)₂], presented good activity against Haemophilus influenzae, a very harmful bacterium to humans.     

Inducible gene expression systems and plant biotechnology

Plant biotechnology relies heavily on the genetic manipulation of crops. Almost invariantly, the gene of interest is expressed in a constitutive fashion, although this may not be strictly necessary for several applications. Currently, there are several regulatable expression systems for the temporal, spatial and quantitative control of transgene activity. These molecular switches are based on components derived from different organisms, which range from viruses to higher eukaryotes. Many inducible systems have been designed for fundamental and applied research and since their initial development, they have become increasingly popular in plant molecular biology.This review covers a broad number of inducible expression systems examining their properties and relevance for plant biotechnology in its various guises, from molecular breeding to pharmaceutical and industrial applications. For each system, we examine some advantages and limitations, also in relation to the strategy on which they rely. Besides being necessary to control useful genes that may negatively affect crop yield and quality, we discuss that inducible systems can be also used to increase public acceptance of GMOs, reducing some of the most common concerns. Finally, we suggest some directions and future developments for their further diffusion in agriculture and biotechnology.     

Quantifying threats in a Mediterranean wetland: are there any changes in their evaluation during a training course?

“Threats analysis” is a conservation discipline directed to identify, quantify, rank, map the human-induced threats occurring in a specific study area, assessing their spatial-temporal regime (e.g., the extension and intensity on specific targets of conservation concern). This approach may be particularly useful in order to define the priorities in the management strategies to be adopted by nature reserves, especially those that are situated in human-altered multi-disturbed contexts. In a remnant Mediterranean wetland of central Italy, where 15 human-induced threats were identified, we compared the assessment given for two regime parameters (scope and severity) of each threat by a panels of not-expert students (n = 13) during different phases of a training academic course, thus evaluating perception modifications during three steps. We applied an experience-based method, then comparing the assessments obtained during the training course with the values obtained previously from a panel of experts (n = 10). Two direct threats (pollution and alien species) significantly changed their mean values in scope during the three steps, and three threats (habitat conversion, garbage, and aircraft) significantly changed as for severity. Students tended to significantly correct their responses in such a way to progressively match with the experts’ responses for a set of threats either for scope (habitat conversion, linear infrastructures, grazing, garbage, fires on reed-beds) or severity (pollution, garbage, fires on reed-beds). Despite the responses were not clear for some threats (aircraft), we suppose that students could assign a initial emphasis towards more charismatic threats as pollution, garbage, fires that instead locally did not appear of priority concern, while threats being uneasily detected (as alien species) were underestimated. Our study showed that a training course with field surveys allows to better define and contextualize the threats affecting a given area, reducing the perception effect that each student had prior to the beginning of the course, when their education information was mainly mass-media based.     

Nitric oxide production inhibited by xenobiotic compounds in the protozoan Paramecium primaurelia

The notable increase in agricultural and industrial activities over the last decades has caused a considerable increase in anthropogenic waste and, consequently, the presence of pollutants in both water and sediments. For this reason, there is great interest in identifying alternative models and bioassays complying with the 3Rs strategy (aimed at Reducing, Refining and Replacing tests on vertebrate organisms in toxicological studies). Protozoa seem to be well suited to this strategy and it is widely accepted that assays with protozoa are relevant to the study of environmental modifications due to the presence of xenobiotic compounds.Recently, we detected the presence of nitric oxide synthase (NOS)-related NADPH-diaphorase activity and neuronal NOS-related molecules, immunologically recognized by the anti-rat brain NOS antibody, in a single-cell freshwater eukaryote, Paramecium primaurelia. In this work we have looked for the basal NO production in living cells of P. primaurelia using the specific fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA) and measuring the intracellular NO levels with image analysis. The NO production was sensitive to compounds modulating NOS activity such as: S-methyl-tiocitrulline, an NOS activity inhibitor, l-NAME, an analogue of arginine that inhibits NO production, arginine, an NOS substrate, or sodium nitroprusside, an NO donor. The NO production in P. primaurelia was also shown to be sensitive to μM concentrations of heavy metals (HgCl₂ and CdCl₂), or μM concentrations of pesticides (diazinon and AFD 25), thus representing a potential biomarker for environmental biomonitoring. The possible involvement of cellular Ca²⁺ concentration, assayed by the fluorescent probe chlortetracycline hydrochloride, in NO production was examined after xenobiotic exposure.     

Amphiphilic porphyrin film on glass as a simple and selective solid-state chemosensor for aqueous Hg2+

Deposition of amphiphilic porphyrin derivatives occurs spontaneously on silanised glass surfaces, in a controlled fashion. The resulting porphyrin films show appreciable fluorescence emission. This emission can be effectively quenched by immersion of the slides into a diluted solution of Hg(2+) (microM concentration). The initial intensity can be restored by washings with a solution of N,N,N',N'-tetrakis(2-pyridilmethyl)ethylenediammine with no loss of efficiency. A remarkable selectivity is featured toward the detection of Hg(2+) over Cu(2+), Cd(2+), Pb(2+) and Zn(2+) counterparts. This protocol can be extended to a flow-through apparatus. The presented results are of importance for the achievement of a solid-state chemosensor for mercuric ions, at micromolar concentration, in water.     

Structure-activity studies on neuropeptide S: identification of the amino acid residues crucial for receptor activation

Neuropeptide S (NPS) has been recently recognized as the endogenous ligand for the previous orphan G-protein-coupled receptor GPR154, now referred to as the NPS receptor (NPSR). The NPS-NPSR receptor system regulates important biological functions such as sleeping/wakening, locomotion, anxiety, and food intake. To collect information on the mechanisms of interaction between NPS and its receptor, a classical structure-activity relationship study was performed. Human (h) NPS derivatives obtained by Ala and d-scan and N- and C-terminal truncation were assessed for their ability to stimulate calcium release in HEK293 cells expressing the human recombinant NPSR. The results of this study indicate that (i) the effect of hNPS is mimicked by the fragment hNPS-(1-10); (ii) Phe(2), Arg(3), and Asn(4) are crucial for biological activity; (iii) the sequence Thr(8)-Gly(9)-Met(10) is important for receptor activation, although with non-stringent chemical requirements; and (iv) the sequence Val(6)-Gly(7) acts as a hinge region between the two above-mentioned domains. However, the stimulatory effect of hNPS given intracerebroventricularly on mouse locomotor activity was not fully mimicked by hNPS-(1-10), suggesting that the C-terminal region of the peptide maintains importance for in vivo activity. In conclusion, this study identified the amino acid residues of this peptide most important for receptor activation.