Lamellipodia and Membrane Blebs Drive Efficient Electrotactic Migration of Rat Walker Carcinosarcoma Cells WC 256

The endogenous electric field (EF) may provide an important signal for directional cell migration during wound healing, embryonic development and cancer metastasis but the mechanism of cell electrotaxis is poorly understood. Additionally, there is no research addressing the question on the difference in electrotactic motility of cells representing various strategies of cell movement—specifically blebbing vs. lamellipodial migration. In the current study we constructed a unique experimental model which allowed for the investigation of electrotactic movement of cells of the same origin but representing different modes of cell migration: weakly adherent, spontaneously blebbing (BC) and lamellipodia forming (LC) WC256 cells. We report that both BC and LC sublines show robust cathodal migration in a physiological EF (1–3 V/cm). The directionality of cell movement was completely reversible upon reversing the field polarity. However, the full reversal of cell direction after the change of EF polarity was much faster in the case of BC (10 minutes) than LC cells (30 minutes). We also investigated the distinct requirements for Rac, Cdc42 and Rho pathways and intracellular Ca²⁺ in electrotaxis of WC256 sublines forming different types of cell protrusions. It was found that Rac1 is required for directional movement of LC to a much greater extent than for BC, but Cdc42 and RhoA are more crucial for BC than for LC cells. The inhibition of ROCK did not affect electrotaxis of LC in contrast to BC cells. The results also showed that intracellular Ca²⁺ is essential only for the electrotactic reaction of BC cells. Moreover, inhibition of MLCK and myosin II did not affect the electrotaxis of LC in contrast to BC cells. In conclusion, our results revealed that both lamellipodia and membrane blebs can efficiently drive electrotactic migration of WC 256 carcinosarcoma cells, however directional migration is mediated by different signalling pathways.     

Thyroid Disorders—Assessments of Trace Elements, Clinical, and Laboratory Parameters

The trace elements studied in this work (Se, Cu, Zn) are the essential constituents or cofactors required to activate numerous enzymes and proteins, playing crucial role in various physiological processes. The disturbed levels of abovementioned elements may adversely affect the endocrine system, resulting in various thyroid disorders among other upsets. The aim of this study was to investigate possible associations between them and parameters of redox balance, thyroid function indices as well as clinical records (duration of disease and therapy, lag time between thyroid surgery and this study examination, LT4 dosage) in patients with different thyroid disorders, including malignant diseases of the gland. In the group of patients with papillary carcinoma, we found a statistically significant higher Cu concentration compared with controls and patients with Hashimoto disease. In the same groups, the parameter of Zn/Cu ratio demonstrated reciprocally arranged statistically significant differences. For the group of papillary cancer patients, there was a negative correlation between lag time since thyroid operation and GPX3 activity. Our data support hypothesis of indirect involvement of Zn and Cu in thyroid regulation. For selenium, lack of simple correlation between its serum level and thyroid indices implies the need for further research on other selenium status parameters more adequately depicting changes in endocrine system.     

The Nucleocapsid Protein of Human Coronavirus NL63

Human coronavirus (HCoV) NL63 was first described in 2004 and is associated with respiratory tract disease of varying severity. At the genetic and structural level, HCoV-NL63 is similar to other members of the Coronavirinae subfamily, especially human coronavirus 229E (HCoV-229E). Detailed analysis, however, reveals several unique features of the pathogen. The coronaviral nucleocapsid protein is abundantly present in infected cells. It is a multi-domain, multi-functional protein important for viral replication and a number of cellular processes. The aim of the present study was to characterize the HCoV-NL63 nucleocapsid protein. Biochemical analyses revealed that the protein shares characteristics with homologous proteins encoded in other coronaviral genomes, with the N-terminal domain responsible for nucleic acid binding and the C-terminal domain involved in protein oligomerization. Surprisingly, analysis of the subcellular localization of the N protein of HCoV-NL63 revealed that, differently than homologous proteins from other coronaviral species except for SARS-CoV, it is not present in the nucleus of infected or transfected cells. Furthermore, no significant alteration in cell cycle progression in cells expressing the protein was observed. This is in stark contrast with results obtained for other coronaviruses, except for the SARS-CoV.     

At Short Telomeres Tel1 Directs Early Replication and Phosphorylates Rif1

The replication time of Saccharomyces cerevisiae telomeres responds to TG_1–3 repeat length, with telomeres of normal length replicating late during S phase and short telomeres replicating early. Here we show that Tel1 kinase, which is recruited to short telomeres, specifies their early replication, because we find a tel1Δ mutant has short telomeres that nonetheless replicate late. Consistent with a role for Tel1 in driving early telomere replication, initiation at a replication origin close to an induced short telomere was reduced in tel1Δ cells, in an S phase blocked by hydroxyurea. The telomeric chromatin component Rif1 mediates late replication of normal telomeres and is a potential substrate of Tel1 phosphorylation, so we tested whether Tel1 directs early replication of short telomeres by inactivating Rif1. A strain lacking both Rif1 and Tel1 behaves like a rif1Δ mutant by replicating its telomeres early, implying that Tel1 can counteract the delaying effect of Rif1 to control telomere replication time. Proteomic analyses reveals that in yku70Δ cells that have short telomeres, Rif1 is phosphorylated at Tel1 consensus sequences (S/TQ sites), with phosphorylation of Serine-1308 being completely dependent on Tel1. Replication timing analysis of a strain mutated at these phosphorylation sites, however, suggested that Tel1-mediated phosphorylation of Rif1 is not the sole mechanism of replication timing control at telomeres. Overall, our results reveal two new functions of Tel1 at shortened telomeres: phosphorylation of Rif1, and specification of early replication by counteracting the Rif1-mediated delay in initiation at nearby replication origins.     

Caucalido platycarpi-Vicietum michauxii — a new weed association from crop fields of Kyrgyzstan (Middle Asia)

The study presents the results of geobotanical investigations conducted in crop fields in the western Tian Shan Mts in Kyrgyzstan (Middle Asia). The main research focused on classification of weed communities developing within this poorly investigated area, were conducted in the vicinity of Bishkek and Kara-Balta in 2010. Altogether, 299 phytosociological relevés were sampled using the Braun-Blanquet method. Based on all segetal vegetation patches, the analyses distinguished a new association: Caucalido platycarpi-Vicietum michauxii. The results of these phytosociological studies fill a gap in the knowledge about the syntaxonomical diversity of the Middle Asia region, which is one of the most crucial for segetal weed species. The study shows that anthropogenic agrocoenoses could harbour relatively rich flora. Extensively cultivated fields could especially serve as a suitable habitat for many xerothermophilous and heliophilous plants. More than 75 species in vegetation plots, mainly permament weeds, have been found. There is also a considerable share of species coming over from swards, screes and meadows.     

Concordant mitochondrial and microsatellite DNA structuring between Polish lowland and Carpathian Mountain wolves

Phylogeographic studies of highly mobile large carnivores suggest that intra-specific genetic differentiation of modern species might be the consequence of the most recent Pleistocene glaciation. However, the relative influence of biogeographical processes and subsequent human-induced population fragmentation requires a better understanding. Poland represents the western edge of relatively continuous distributions of many wide-ranging species, e.g. lynx (Lynx lynx), wolves (Canis lupus), moose (Alces alces) and, therefore, a key area for understanding historic and contemporary patterns of gene flow in central Europe. We examined wolf genetic structure in Poland and in a recently recolonized area in eastern Germany using microsatellite profiles (n = 457) and mitochondrial DNA sequencing (mtDNA, n = 333) from faecal samples. We found significant genetic structure and high levels of differentiation between wolves in the Carpathian Mountains and the Polish lowlands. Our findings are consistent with previously reported mtDNA subdivision between northern lowlands and southern mountains, and add new and concordant findings based on autosomal marker variation. Wolves in western Poland and eastern Germany showed limited differentiation from northeastern Poland. Although the presence of private alleles suggests immigration also from areas not sampled in this study, most individuals seem to be immigrants from northeastern Poland or their descendants. We observed moderate genetic differentiation between certain northeastern lowland regions separated by less than 50 km. Moreover, mtDNA results indicated a southeastern subpopulation near the border with Ukraine. The observed structure might reflect landscape fragmentation and/or ecological differences resulting in natal habitat-biased dispersal.     

Synthesis,biological activity and solution structure of new analogues of the antimicrobial Gramicidin S

Gramicidin S (GS) is a cyclo‐decapeptide antibiotic isolated from Bacillus brevis. The structural studies have shown that GS forms a two‐stranded antiparallel β‐sheet imposed by two II′ β‐turns. Despite its wide Gram+ and Gram? antimicrobial spectrum, GS is useless in therapy because of its high hemotoxicity in humans. It was found, however, that the analogues of GS‐14 (GS with 14 amino acid residues) attained a better antimicrobial selectivity when their amphipatic moments were perturbed. In this study, we report effects of similar perturbations imposed on GS cyclo‐decapeptide analogues. Having solved their structures by NMR/molecular dynamics and having tested their activities/selectivities, we have concluded that the idea of perturbation of the amphipatic moment does not work for GS‐10_0 analogues. An innovative approach to the synthesis of head‐to‐tail cyclopeptides was used. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd.     

Induction of apoptosis in the grain aphid Sitobion avenae (Hemiptera: Aphididae) under the influence of phytohaemagglutinin PHA

In the current work, the toxicity and mechanism of phytohaemagglutinin (PHA), lectin isolated from the kidney bean (Phaseolus vulgaris) to the grain aphid (Sitobion avenae F.) were studied. When S. avenae was fed an artificial diet containing the lectin PHA, toxicity assays indicated that fecundity decreased, the pre-reproductive period and generation time were prolonged, and mortality increased. To elucidate the mode of action of PHA, the interaction between the lectin and the insect gut was investigated. Interestingly, DNA fragmentation was observed in extract of gut of treated grain aphid, and this was accompanied by an increase in caspase-3 activity. Results indicate that the detrimental activity of PHA on S. avenae may involve effects on death of the gut epithelial cells.     

Toxic and deterrent effects of phytohemagglutinin on the grain aphid Sitobion avenae

The toxicity and deterrent effects of phytoheamagglutinin (Phaseolus vulgaris agglutinin, PHA) to the grain aphid (Sitobion avenae F.) was investigated in the laboratory by use of liquid diets or gels containing various PHA concentrations. The addition of PHA to the liquid diet increased the aphid pre-reproductive period, mortality, and generation time and decreased aphid fecundity and intrinsic rate of natural increase. Aphids fed on sucrose-agarose gels with higher concentrations of PHA exhibited no phloem-feeding behavior. The number of gel penetrations was reduced and duration of the pathway phase was increased as PHA concentration increased. These results indicate that PHA is detrimental to S. avenae and that the effect of PHA is at least partially explained by suppression of feeding behaviour. PHA could be useful for managing S. avenae.