Design,synthesis, in silico studies and in vitro evaluation of isatin-pyridine oximes hybrids as novel acetylcholinesterase reactivators

Organophosphorus poisoning caused by some pesticides and nerve agents is a life-threating condition that must be swiftly addressed to avoid casualties. Despite the availability of medical countermeasures, the clinically available compounds lack a broad spectrum, are not effective towards all organophosphorus toxins, and have poor pharmacokinetics properties to allow them crossing the blood-brain barrier, hampering cholinesterase reactivation at the central nervous system. In this work, we designed and synthesised novel isatin derivatives, linked to a pyridinium 4-oxime moiety by an alkyl chain with improved calculated properties, and tested their reactivation potency against paraoxon- and NEMP-inhibited acetylcholinesterase in comparison to the standard antidote pralidoxime. Our results showed that these compounds displayed comparable in vitro reactivation also pointed by the in silico studies, suggesting that they are promising compounds to tackle organophosphorus poisoning.     

The use of oxytocin in liquid semen doses to reduce seasonal fluctuations in the reproductive performance of sows and improve litter parameters—a 2-year study

The objective of the present research was to eliminate seasonal fluctuations in year-round reproductive performance of sows and to improve litter parameters by administration of oxytocin into liquid semen insemination doses. A 2-year experiment was performed on crossbreed sows, Polish Large White × Polish Landrace, which were partitioned into two groups: control, insemination without any modification with 100 mL semen doses and oxytocin, insemination with 100 mL semen doses to which 5 IU of oxytocin was added just before insemination. A total of 10,486 inseminations were made. The farrowing rate and obtained litter parameters, including the effect of season, were analyzed. For each litter, the following factors were defined: average litter size, percentage of fetal death and mummified piglets, average piglet birth weight, percentage of piglet mortality, fecundity index, average number of piglets weaned, weaned piglet weight, and daily gain. Sows presented a positive reaction to the experimental factor. A statistically higher farrowing rate for oxytocin group in summer and autumn seasons was confirmed (P ≤ 0.01). Regardless of the season, a higher average litter size was observed in the oxytocin group with the most evident differences for winter, spring (P ≤ 0.01), and summer (P ≤ 0.05). The effect of oxytocin on the percentage of fetal death and mummified piglets born was not confirmed statistically except for winter. Analyzing the fecundity index, higher values were obtained for the oxytocin group in all seasons (P ≤ 0.01), including the lowest difference between groups for winter (51.43) and the highest for summer (100.61). A higher average birth piglet weight and weaned piglet weight were recorded for the oxytocin group in all seasons. The highest differences in birth piglet weight between groups were noted for spring (0.22 kg; P ≤ 0.01) and winter (0.17 kg; P ≤ 0.05) and in weaned piglet weight for winter and spring (0.58 kg and 0.52 kg; for both, P ≤ 0.01). The greatest daily gains were observed in the winter season (P ≤ 0.05) in favor of oxytocin. On the basis of the presented results, it should be noted that the use of oxytocin into insemination doses improves the farrowing rate and other parameters of the reproductive performance of sows. In the absence of negative effects, year-round insemination with oxytocin addition into seminal doses is recommended, which effectively improves the production performance and reduces the problem of seasonality in reproduction.     

Palaeovegetation in the Pavlovské vrchy hills region (South Moravia,Czech Republic) around 25,000 bp: the Bulhary core

This paper presents results of pollen and macroscopic analyses of a geological core from the village of Bulhary, at the north-eastern foot of the limestone Pavlovské vrchy hills, South Moravia, Czech Republic. The core reached down to about 10 m of aeolic loess layers and some 40–50 cm of compressed organogenic sediments. They accumulated around ca 25,000 years bp. The moss peat and algal gyttja we have analysed were deposited in an ox-bow of the past flood-plain of the river Dyje. The time of the sedimentation may be synchronous with the time when the northern foot of the Pavlovské vrchy hills was inhabited by upper Palaeolithic people of the Gravettian (Pavlovian) culture, dated generally to an Upper Würmian interstadial. Archaeologists excavated their settlements in the villages of Dolní Věstonice and Pavlov, neighbouring our site of Bulhary. Our analyses permit the reconstruction of the vegetation of that time in the region but may also reveal the environment and living conditions of the Palaeolithic people. In our assemblages over 200 types of palynomorphs (pollen, spores, etc.) have been found, which indicates a very rich flora and vegetation growing in at least six different biotopes of the Pavlovské vrchy hills region: (1) Open, mostly coniferous stands with grass and herbaceous cover in their undergrowth. These were probably situated in the lowest parts of the territory and could also have dominated in the broader vicinity with loess cover. (2) Grass and herbaceous steppe-like vegetation probably occupied the belt above where the trees occurred but still within the loess zone. (3) The highest elevations of the hills and their limestone cliffs formed biotopes for a complex of subalpine (alpine) communities. (4) The flood plain was certainly occupied by very diversified tall-herb vegetation with scattered alder and willow trees or shrubs. (5) Aquatic communities of the infilled stages in the river ox-bows are well documented by microscopic and macroremain finds of water macrophytes, algae and mosses. (6) The presence of scattered spring outflows and their communities is indicated by pollen of typical spring plants, e.g. Chrysosplenium, Montia, Cardamine amara etc. The vegetation, especially that in the flood-plain, produced enough plant biomass to feed numerous herds of large herbivorous mammals, which were hunted by the Palaeolithic tribes. Also the climate was probably relatively suitable for the style of living at that time. It could have been similar to that of the present northern boreal zone of NE Scandinavia and N Russia or even milder.     

The RNase H-like superfamily: new members,comparative structural analysis and evolutionary classification

Ribonuclease H-like (RNHL) superfamily, also called the retroviral integrase superfamily, groups together numerous enzymes involved in nucleic acid metabolism and implicated in many biological processes, including replication, homologous recombination, DNA repair, transposition and RNA interference. The RNHL superfamily proteins show extensive divergence of sequences and structures. We conducted database searches to identify members of the RNHL superfamily (including those previously unknown), yielding >60 000 unique domain sequences. Our analysis led to the identification of new RNHL superfamily members, such as RRXRR (PF14239), DUF460 (PF04312, COG2433), DUF3010 (PF11215), DUF429 (PF04250 and COG2410, COG4328, COG4923), DUF1092 (PF06485), COG5558, OrfB_IS605 (PF01385, COG0675) and Peptidase_A17 (PF05380). Based on the clustering analysis we grouped all identified RNHL domain sequences into 152 families. Phylogenetic studies revealed relationships between these families, and suggested a possible history of the evolution of RNHL fold and its active site. Our results revealed clear division of the RNHL superfamily into exonucleases and endonucleases. Structural analyses of features characteristic for particular groups revealed a correlation between the orientation of the C-terminal helix with the exonuclease/endonuclease function and the architecture of the active site. Our analysis provides a comprehensive picture of sequence-structure-function relationships in the RNHL superfamily that may guide functional studies of the previously uncharacterized protein families.     

Trepostome bryozoans from the Zahořany Formation (Upper Ordovician) of Loděnice,Prague Basin,Czech Republic

Three trepostome bryozoan species are described from the Upper Ordovician Zaho?any Formation of Loděnice, Prague Basin, Czech Republic. One genus is new—Lodenicella gen. nov. One species is described in open nomenclature. The described fauna contains ramose colonies or ramose branched projections from encrusting tubular-shaped colonies which inhabited shallow environment with moderate wave energy and significant influx of clastic material.     

Dielectric properties of two diastereoisomers of the arabinose and their equimolar mixture

Dielectric relaxation measurements were performed on two enantiomers, d- and l-arabinose and their equimolar mixture, and compared to dielectric data obtained for d-ribose. d-Arabinose differs from d-ribose by having the opposite configuration at C2. This study reveals that both d- and l- of arabinose exhibit α-relaxation peaks with the same shape for the same α-relaxation time τ_α, and the same steepness index for the T_g-scale T-dependence of τ_α. However, the two isomers have slightly different glass transition temperatures T_g’s, and their secondary γ-relaxation times also differ slightly from the previously observed γ-relaxation in d-ribose at the same temperature. However, when samples of both investigated monosaccharides are annealed at higher temperatures, their glass transition temperatures become nearly identical. This is an effect of the mutarotation process, which leads to the formation of pairs of the enantiomers and accordingly they should have the same physical properties. The width of the α-relaxation of d- and l-arabinose is broader than that of d-ribose, as reflected by the smaller stretch exponent in the Kohlrausch-Williams-Watts function used to fit the data of the former (β_KWW = 0.46 ± 0.01) than the latter (β_KWW = 0.55 ± 0.01). The width of the α-relaxation of racemic mixture of the d- and l-arabinose is slightly broader than that of the pure isomers. While the dielectric loss data of d-ribose in the glassy state at ambient and elevated pressures show an inflexion indicating the presence of the JG β-relaxation, the data of d- and l-arabinose show no such feature for identification of the supposedly universal JG β-relaxation. Nevertheless, on comparing the loss spectra of d-arabinose with that of d-ribose, the presence of the JG β-relaxation in d-arabinose has been rationalized.     

The evolution of an 'intelligent' dispersal strategy: biased, correlated random walks in patchy landscapes

Theoretical work exploring dispersal evolution focuses on the emigration rate of individuals and typically assumes that movement occurs either at random to any other patch or to one of the nearest‐neighbour patches. There is a lack of work exploring the process by which individuals move between patches, and how this process evolves. This is of concern because any organism that can exert control over dispersal direction can potentially evolve efficiencies in locating patches, and the process by which individuals find new patches will potentially have major effects on metapopulation dynamics and gene flow. Here, we take an initial step towards filling this knowledge gap. To do this we constructed a continuous space population model, in which individuals each carry heritable trait values that specify the characteristics of the biased correlated random walk they use to disperse from their natal patch. We explore how the evolution of the random walk depends upon the cost of dispersal, the density of patches in the landscape, and the emigration rate. The clearest result is that highly correlated walks always evolved (individuals tended to disperse in relatively straight lines from their natal patch), reflecting the efficiency of straight‐line movement. In our models, more costly dispersal resulted in walks with higher correlation between successive steps. However, the exact walk that evolved also depended upon the density of suitable habitat patches, with low density habitat evolving more biased walks (individuals which orient towards suitable habitat at quite large distances from that habitat). Thus, low density habitat will tend to develop individuals which disperse efficiently between adjacent habitat patches but which only rarely disperse to more distant patches; a result that has clear implications for metapopulation theory. Hence, an understanding of the movement behaviour of dispersing individuals is critical for robust long‐term predictions of population dynamics in fragmented landscapes.     

Tests with Daphnia magna: a new approach to prescreen toxicity of newly synthesized acetylcholinesterase reactivators

Reactivators of phosphorylated acetylcholinesterase (oximes) are substances used as a human antidotal therapy for organophosphate poisoning. The objective of our study was to examine if juveniles of freshwater microcrustacean Daphnia magna could be employed as test animals in early screen toxicity tests of those substances as a first step for further experiments with daphnids intoxicated by organophosphates. For this purpose, seven different oximes were investigated. It was found that toxicity of all tested oximes increased with time. Mono-quaternary oximes were approximately ten fold (EC50, 14.9 mg.l(-1)) more toxic in 24 hour tests and five fold (EC50 was 79.46 mg.l(-1)) more toxic in 48 hour tests than bis-quaternary oximes. Tests with daphnids were shown to be easy to carry out at low cost and provided valuable results which could be used as a starting point for further research.