Development of 5-fluorouracil-dichloroacetate mutual prodrugs as anticancer agents.

5-Fluorouracil (5-FU) is one of the most widely applied chemotherapeutic agents with a broad spectrum of activity. However, despite this versatile activity, its use poses many limitations. Herein, novel derivatives of 5-FU and dichloroacetic acid have been designed and synthesized as a new type of codrugs, also known as mutual prodrugs, to overcome the drawbacks of 5-FU and enhance its therapeutic efficiency. The stability of the obtained compounds has been tested at various pH values using different analytical techniques, namely HPLC and potentiometry. The antiproliferative activity of the new 5-FU derivatives was assessed in vitro on SK-MEL-28 and WM793 human melanoma cell lines in 2D culture as well as on A549 human lung carcinoma, MDA-MB-231 breast adenocarcinoma, LL24 normal lung tissue, and HMF normal breast tissue as a multicellular 3D spheroid model cultured in standard (static) conditions and with the use of microfluidic systems, which to a great extent resembles the in vivo environment. In all cases, new mutual prodrugs showed a higher cytotoxic activity toward cancer models and lower to normal cell models than the parent 5-FU itself.

Mites inhabiting nests of wood warbler, Phylloscopus sibilatrix (Aves: Passeriformes), in the Wielkopolska National Park in western Poland.

The wood warbler, Phylloscopus sibilatrix (Aves: Passeriformes), is a well-known model organism for studying bird migration, breeding habitat selection and nest predation. The nest acarofauna of this bird species has not been extensively studied so far. To provide a comprehensive report on mite species inhabiting wood warbler nests and to assess infestation parameters (prevalence, intensity, and abundance) for mite species and orders, we collected 45 nests of this bird species in the Wielkopolska National Park in western Poland. Analyses revealed a huge diversity (198 species) of mites inhabiting wood warbler nests. We found individuals belonging to the Mesostigmata, Trombidiformes and Sarcoptiformes. The Trombidiformes, represented in our study only by the Prostigmata, achieved statistically significantly lower intensity and abundance, compared to representatives of other orders. However, the number of recorded prostigmatid species was high (65). The most common were: Stigmaeus sphagneti (22 nests), Stigmaeus longipilis (16), Eupodes voxencollinus (15), Cunaxa setirostris (14), Stigmaeus pilatus (11), and Linopodes sp. 2 (10). The prevalence of Mesostigmata and Sarcoptiformes was equal, reaching 91.1%. Most of Gamasina (Mesostigmata) species found in this study were more characteristic of the soil environment and forest litter than bird nests, but there was also a typical bird parasite, viz. Ornithonyssus sylviarum. None of the observed species of Uropodina (Mesostigmata) or Oribatida (Sarcoptiformes) was typical for bird nests. Among the Uropodina, the highest parameters of nest infestation were achieved by Oodinychus ovalis, whereas among the Oribatida, they were achieved by Metabelba pulverosa. We discuss the importance of wood warbler nests for mite dispersal, survival and reproduction.

Effective specialist or jack of all trades? Experimental evolution of a crop pest in fluctuating and stable environments.

Understanding pest evolution in agricultural systems is crucial for developing effective and innovative pest control strategies. Types of cultivation, such as crop monocultures versus polycultures or crop rotation, may act as a selective pressure on pests' capability to exploit the host's resources. In this study, we examined the herbivorous mite Aceria tosichella (commonly known as wheat curl mite), a widespread wheat pest, to understand how fluctuating versus stable environments influence its niche breadth and ability to utilize different host plant species. We subjected a wheat-bred mite population to replicated experimental evolution in a single-host environment (either wheat or barley), or in an alternation between these two plant species every three mite generations. Next, we tested the fitness of these evolving populations on wheat, barley, and on two other plant species not encountered during experimental evolution, namely rye and smooth brome. Our results revealed that the niche breadth of A. tosichella evolved in response to the level of environmental variability. The fluctuating environment expanded the niche breadth by increasing the mite's ability to utilize different plant species, including novel ones. Such an environment may thus promote flexible host-use generalist phenotypes. However, the niche expansion resulted in some costs expressed as reduced performances on both wheat and barley as compared to specialists. Stable host environments led to specialized phenotypes. The population that evolved in a constant environment consisting of barley increased its fitness on barley without the cost of utilizing wheat. However, the population evolving on wheat did not significantly increase its fitness on wheat, but decreased its performance on barley. Altogether, our results indicated that, depending on the degree of environmental heterogeneity, agricultural systems create different conditions that influence pests' niche breadth evolution, which may in turn affect the ability of pests to persist in such systems.

Mechanisms of dispersal and colonisation in a wind-borne cereal pest, the haplodiploid wheat curl mite.

Dispersal and colonisation determine the survival and success of organisms, and influence the structure and dynamics of communities and ecosystems in space and time. Both affect the gene flow between populations, ensuring sufficient level of genetic variation and improving adaptation abilities. In haplodiploids, such as Aceria tosichella (wheat curl mite, WCM), a population may be founded even by a single unfertilised female, so there is a risk of heterozygosity loss (i.e. founder effect). It may lead to adverse outcomes, such as inbreeding depression. Yet, the strength of the founder effect partly depends on the genetic variation of the parental population. WCM is an economically important pest with a great invasive potential, but its dispersal and colonisation mechanisms were poorly studied before. Therefore, here we assessed WCM dispersal and colonisation potential in relation to the genetic variation of the parental population. We checked whether this potential may be linked to specific pre-dispersal actions (e.g. mating before dispersal and collective behaviour). Our study confirms that dispersal strategies of WCM are not dependent on heterozygosity in the parental population, and the efficient dispersal of this species depends on collective movement of fertilised females.

A sink host allows a specialist herbivore to persist in a seasonal source.

In seasonal environments, sinks that are more persistent than sources may serve as temporal stepping stones for specialists. However, this possibility has to our knowledge, not been demonstrated to date, as such environments are thought to select for generalists, and the role of sinks, both in the field and in the laboratory, is difficult to document. Here, we used laboratory experiments to show that herbivorous arthropods associated with seasonally absent main (source) habitats can endure on a suboptimal (sink) host for several generations, albeit with a negative growth rate. Additionally, they dispersed towards this host less often than towards the main host and accepted it less often than the main host. Finally, repeated experimental evolution attempts revealed no adaptation to the suboptimal host. Nevertheless, field observations showed that arthropods are found in suboptimal habitats when the main habitat is unavailable. Together, these results show that evolutionary rescue in the suboptimal habitat is not possible. Instead, the sink habitat functions as a temporal stepping stone, allowing for the persistence of a specialist when the source habitat is gone.

A Preliminary Assessment of Amblyseius andersoni (Chant) as a Potential Biocontrol Agent against Phytophagous Mites Occurring on Coniferous Plants.

Development, survival and reproduction of Ambyseius andersoni (Chant), a predatory mite widely distributed in Europe, were assessed on different food items. These included two key pests of ornamental coniferous plants, i.e., Oligonychus ununguis (Jacobi) and Pentamerismus taxi (Haller) and pollen of Pinus sylvestris L. The rationale behind these experiments was to provide a preliminary assessment of the potential of A. andersoni as a biocontrol agent of the above phytophagous arthropods and evaluate pine pollen as an alternative food source for the predator. Under laboratory conditions (23 ± 0.5 °C, 70 ± 10% RH and 16L:8D) A. andersoni was able to feed, develop and reproduce on all tested diets. The shortest development time (egg to female) was obtained when the predator fed on P. taxi (mean = 5.12 d) and the longest was on pine pollen (mean = 6.55 d). The rm value was significantly higher on both tested prey (0.166 on P. taxi and 0.160 on O. ununguis) than on pollen (0.139). Thus, we do not recommend pine pollen for mass rearing of A. andersoni; however, we conclude that pollen may provide sufficient sustenance for the predator population under field conditions when prey are absent. The potential of A. andersoni as a biocontrol agent of O. ununguis and P. taxi is discussed.

New insight into nucleo α-amino acids - Synthesis and SAR studies on cytotoxic activity of ß-pyrimidine alanines.

Three series of the ß-pyrimidine alanines, including willardiine - a naturally occurring amino acid, were prepared from the l-serine-derived sulfamidates. Compounds 3b, 4a and 4b demonstrated antiproliferative activity toward the studied cancer cell lines, albeit the effect of these compounds on human brain astrocytoma MOG-G-CCM cells was more significant than on human neuroblastoma SK-N-AS cells. The cytosine analog of willardiine, compound 4b, reduced viability of MOG-G-CCM cells with EC50 = 36 ± 2 µM, more effectively than AMPA antagonist GYKI 52466. Willardiine showed possible capability of affecting invasiveness of glioblastoma U251 MG cells with no effect on their viability and morphology. Compound 3d, the ethyl ester of willardiine, featured activity toward binding domain hHS1S2I of the GluR2 receptor. Docking analysis revealed that the location mode of compound 3d at the S1S2 domain of hGluR2 (PDB ID: 3R7X) might differ from that of willardiine.

A novel experimental approach for studying life-history traits of phytophagous arthropods utilizing an artificial culture medium.

Experimental approaches to studying life-history traits in minute herbivorous arthropods are hampered by the need to work with detached host plant material and the difficulty of maintaining that material in a suitable condition to support the animal throughout the duration of the test. In order to address this shortcoming, we developed a customizable agar-based medium modified from an established plant cell-culture medium to nourish detached leaves laid atop it while also preventing arthropods from escaping the experimental arena. The artificial culture medium was tested with two herbivorous mite species: the wheat curl mite (Aceria tosichella; Eriophyidae) and two-spotted spider mite (Tetranychus urticae; Tetranychidae). The proposed approach was a major improvement over a standard protocol for prolonged studies of individual eriophyid mites and also provided some benefits for experiments with spider mites. Moreover, the described method can be easily modified according to the requirements of host plant species and applied to a wide range of microherbivore species. Such applications include investigations of life-history traits and other ecological and evolutionary questions, e.g. mating or competitive behaviours or interspecific interactions, assessing invasiveness potential and predicting possible outbreaks. The approach presented here should have a significant impact on the advancement of evolutionary and ecological research on microscopic herbivores.

Potential bioisosteres of ß-uracilalanines derived from 1H-1,2,3-triazole-C-carboxylic acids.

The 1H-1,2,3-triazole-originated derivatives of willardiine were obtained by: (i) construction of the 1H-1,2,3-triazole ring in 1,3-dipolar cycloaddition of the uracil-derived azides and the carboxylate-bearing alkynes or α-acylphosphorus ylide, or (ii) N-alkylation of the uracil derivative with the 1H-1,2,3-triazole-4-carboxylate-derived mesylate. The latter method offered: (i) reproducible results, (ii) a significant reduction of amounts of auxiliary materials, (iii) reduction in wastes and (iv) reduction in a number of manual operations required for obtaining the reaction product. Compound 6a exhibited significant binding affinity to hHS1S2I ligand-binding domain of GluR2 receptor (EC50 = 2.90 µM) and decreased viability of human astrocytoma MOG-G-CCM cells in higher extent than known AMPA antagonist GYKI 52466.

Typhlodromus pyri and Euseius finlandicus (Acari: Phytoseiidae) as potential biocontrol agents against spider mites (Acari: Tetranychidae) inhabiting willows: laboratory studies on predator development and reproduction on four diets.

Typhlodromus pyri Scheuten and Euseius finlandicus (Oudemans) are important predators of phytophagous mites. The present laboratory study aimed to determine whether both species can develop and reach maturity feeding on spider mites occurring on willows, i.e., Schizotetranychus schizopus (Zacher), Schizotetranychus garmani Pritchard & Baker, and Tetranychus urticae Koch, and on Brassica napus L. pollen. The predators' development, reproduction and demographic parameters were significantly affected by diet. The data suggest that rape pollen can be useful in mass rearing of E. finlandicus but is completely unsuitable as alternative food for T. pyri. Short development time and high values of population parameters achieved by T. pyri feeding on larvae and protonymphs of S. schizopus and by E. finlandicus feeding on juvenile stages of S. garmani indicate great suitability of these preys as food for the phytoseiids, and make both predatory species promising biocontrol agents in spider mite control on willows.

Synthesis of novel aza-analogues of tiazofurin with 2-[5,5-bis(hydroxymethyl)pyrrolidin-2-yl] framework as sugar mimic.

The novel aza-analogues of tiazofurin (TZF) with 2-[5,5-bis(hydroxymethyl)pyrrolidin-2-yl] moiety, as sugar mimic, were synthesized from O,O-cyclohexylidene derivative of 4,4-bis(hydroxymethyl)-4-nitrobutanal in multi-gram scale. The synthetic route consisted of three stages: (i) the synthesis of corresponding derivative of 5,5-bis(hydroxymethyl)pyrrolidine-2-carbonitrile, (ii) the construction of ethyl thiazole-2-carboxylate part by the conversion of the pyrrolidine-2-carbonitrile into the N-trifluoroacetyl derivative followed by cyclocondensation with L-cysteine ethyl ester and then by dehydrogenation, and (iii) the final transformation of the ethyl thiazole-4-carboxylate into the aza-analogues of TZF. The TZF aza-analogues were evaluated for their antiviral activities in cell-culture-based assays.

Complexation of metal ions in Langmuir films formed with two amphiphilic dioxadithia crown ethers.

The two new crown ethers presented in this study were synthesized in order to investigate two important features of ionophores, namely metal cation complexation and interfacial properties, and the way in which they interrelate. The two derivatives were conceived as analogs of membrane phospholipids with respect to their amphiphilicity and geometry. They contain a hydrophilic 1,1'-dioxo-3,3'-dithio-14-crown ether headgroup and bear two myristoyl or stearoyl lateral chains. The length of the myristoyl and stearoyl derivatives in an extended conformation is comparable with the thickness of the individual leaflets of cell membranes. The membrane-related and complexation properties of the two crown ether derivatives were studied in monomolecular films spread on pure water and on aqueous solutions of mono-, di-, and trivalent metal salts. The properties of the monolayers are described quantitatively using thermodynamic models. The compression isotherms of the monolayers formed on different subphases show a clear-cut differentiation of the monovalent and di- or trivalent cations with both ligands. This differentiation was interpreted in terms of conformational changes occurring in the crown ether derivatives upon complexation. Molecular modeling indicates that the mono- and divalent cations are coordinated differently by the ligands, yielding complexes with different conformations. The differences of the conformations of the mono- and di- or trivalent cation complexes may be important from the point of view of the interactions with lipid membranes and the biological activity of these potential ionophores.