Abrupt shift from clear to turbid state in a shallow eutrophic, biomanipulated lake

Monitoring data were used to assess causes behind a recent shift from a clear-water to a turbid-water state in Lake Major, a 10 ha shallow lake in Hungary. In 1999–2000, fish manipulation was conducted in this hypertrophic lake. Reduced fish stock resulted in clearing water and the development of a dense (>80% coverage) submerged vegetation in 2005. During the recent abrupt shift, which occurred in 2007, submerged vegetation subsequently declined after a two-year period of clear water and abundant vegetation. An intense decay of macrophytes within the lake produced a rapid transition between the clear- and turbid-water states. During the clear-water state in 2005–2006, the most important variables predominantly correlating with macrophyte cover were Secchi transparency, temperature and TN, while TN, temperature, Secchi depth and chlorophyll-a were the most significant variables during the turbid-water state in 2007. Nitrogen may play a significant role in the cover of submerged macrophytes when TP is moderate. We argue that several factors in concert are necessary to initiate a shift. Water temperature likely has contributed to triggering shift through inter-year-dependent changes in cover of macrophytes, with fish recruitment having key roles in the dynamics of shallow lakes. Handling editor: Luigi Naselli-Flores     

Olfactory response of predatory mite Phytoseiulus persimilis to synthetic methyl salicylate

We studied the role of methyl salicylate as an important part of volatile blends in searching behaviour of the predatory mite, Phytoseiulus persimilis (Athias-Henriot). We showed that Methyl Salicylate (MeSa) attracted the predatory mites (1 h starved) in a dose range of 0.02–20?μg. The highest attraction was recorded at the dose of 0.2?μg. The dose of 0.002?μg was considered under the detection level of the predator. P. persimilis could not detect the extra amount of MeSa (100 and 200?μg). The probable effect of starvation period on the predator response is discussed.     

Cell cycle-dependent chromatin shuttling of HBO1–JADE1 histone acetyl transferase (HAT) complex

HAT HBO1 interacts with 2 isoforms of JADE1: JADE1S and JADE1L. JADE1 promotes acetylation of nucleosomal histones by HBO1. HBO1–JADE1 complex facilitates cell proliferation by unclear mechanisms. Here we report intracellular chromatin shuttling of HBO1–JADE1 complex during mitosis coupled to phosphorylation of JADE1. In interphase of dividing cells JADE1S was localized to the nucleus and associated with chromatin. As cells approached mitosis, specifically prophase, JADE1S dissociated from chromatin and associated with cytoplasm. JADE1S chromatin re-association began in telophase and paralleled nuclear envelope membrane reassembly. By early G₁, JADE1S was re-associated with chromatin and localized to the nucleus. Importantly, cytoplasmic but not chromatin-associated JADE1 protein was phosphorylated. Mass-Spectrometric analysis of JADE1S protein isolated from G₂/M-arrested cells identified 6 phosphorylated amino acid residues: S89, T92, S102, S121, S392, and T468, including 3 novel sites. Temporally, JADE1S phosphorylation and dephosphorylation during mitosis correlated with JADE1S chromatin dissociation and recruitment. JADE1S chromatin recruitment was accompanied by the global histone H4 acetylation. Pharmacological inhibitor of Aurora A kinase prevented JADE1S protein band shift and chromatin dissociation, suggesting regulatory function for phosphorylation. In vivo experiments supported our in vitro results. In mouse kidneys, JADE1S transiently accumulated in the cytoplasm of tubular epithelial cells during kidney regeneration. The transient increase in the number of cells with cytoplasmic JADE1S directly correlated with activation of tubular cell proliferation and inversely correlated with the number of cells with nuclear JADE1S staining, supporting biological role of HBO1–JADE1 shuttling during organ regeneration.     

Historic range dynamics in Kaiser's mountain newt (Neurergus kaiseri): Insights from phylogeographic analyses and species distribution modeling

Vulnerable Kaiser''s mountain newt, Neurergus kaiseri, is endemic to highland streams, springs, and pools of the southwestern Zagros mountain, Iran. The present study aimed to use an integration of phylogeographical and species distribution modeling (SDM) approaches to provide new insights into the evolutionary history of the species throughout Quaternary climate oscillations. The phylogeographical analysis was followed by analyzing two mitochondrial DNA (mt‐DNA) markers including 127 control region (D‐loop) and 72 NADH dehydrogenase 2 (ND2) sequences from 15 populations in the entire species range that were obtained from GenBank. Potential recent and past distribution (the Last Glacial Maximum, LGM, 21 Kya and the Mid‐Holocene, 6 Kya) reconstructed by ensemble SDM using nine algorithms with CCSM4, MIROC‐ESM, and MPI‐ESM‐P models. N. kaiseri displayed two distinct lineages in the northern and southern regions that diverged in the Early‐Pleistocene. The demographics analysis showed signs of a slight increase in effective population size for both northern and southern populations in the Mid‐Pleistocene. Biogeography analysis showed that both vicariance and dispersal events played an important role in the formation of recent species distribution of N. kaiseri. Based on SDM projection onto paleoclimatic data, N. kaiseri displayed a scenario of past range expansion that followed by postglacial contraction. The models showed that the distribution range of the species may have shifted to a lower altitude during LGM while with amelioration of climatic during Mid‐Holocene to recent conditions caused the species to shift to the higher altitude. The findings of the current study support the hypothesis that the Zagros mountains​ may be acting as climatic refugia and play an important role in the protection of isolated populations during climate oscillations.     

Mitochondrial DNA sequence analysis reveals multiple Pleistocene glacial refugia for the Yellow‐spotted mountain newt,Neurergus derjugini (Caudata: Salamandridae) in the mid‐Zagros range in Iran and Iraq

Phylogeography is often used to investigate the effects of glacial cycles on current genetic structure of various plant and animal species. This approach can also identify the number and location of glacial refugia as well as the recolonization routes from those refugia to the current locations. To identify the location of glacial refugia of the Yellow‐spotted mountain newt, Neurergus derjugini, we employed phylogeography patterns and genetic variability of this species by analyzing partial ND4 sequences (867 bp) of 67 specimens from 15 sampling localities from the whole species range in Iran and Iraq. Phylogenetic trees concordant with haplotype networks showed a clear genetic structure among populations as three groups corresponding to the populations in the north, center, and south. Evolutionary ages of clades north and south ranging from 0.15 to 0.17 Myr, while the oldest clade is the central clade, corresponding to 0.32 Myr. Bayesian skyline plots of population size change through time show a relatively slight increase until about 25 kyr (around the last glacial maximum) and a decline of population size about 2.5 kyr. The presence of geographically structured clades in north, center, and south sections of the species range signifies the disjunct populations that have emerged in three different refugium. This study illustrates the importance of the effect of previous glacial cycles in shaping the genetic structure of mountain species in the Zagros range. These areas are important in terms of long‐term species persistence and therefore valuable areas for conservation of biodiversity.     

Development of Transgenic Mice Expressing Calcitonin as a Beta-lactoglobulin Fusion Protein in Mammary Gland

Expression of foreign proteins in mammalian milk is becoming a widespread strategy for high-level production of recombinant pharmaceuticals, especially those with the most complex post-translational modifications. A milk-specific ovine beta-lactoglobulin (oBLG) promoter was used to drive expression of recombinant calcitonin in mouse milk. A gene construct was generated, consisting of 10.7 kbp of the oBLG gene including its promoter and 3′ flanking region with the calcitonin coding sequences inserted in-frame into the oBLG fifth exon. After microinjection, six founder mice transmitted the transgene to their progeny. RT-PCR confirmed mammary-gland specific expression of recombinant mRNA in most transgenic mice and Western blot analysis confirmed expression of chimeric protein. Calcitonin can thus be expressed under the oBLG promoter and regulatory elements in a mammary-gland specific manner.     

Small area inequalities in breast cancer mortality and screening

Despite the unfavourable epidemiological status, the Hungarian breast cancer control is a non-appropriately developed system having considerable geographical inequalities. The study objective was to describe the small-area pattern of breast cancer mortality and of frequency of mammographical examination. The influence of socio-economical status on these patterns was also studied. The standardised mortality ratios and the standardised relative frequency of mammography was determined for settlements, zipcode areas and small regions. Their correlations were analysed with education, unemployment ratio, ratio of Gypsy and German ethnic minorities, population size, smoking, distance to the nearest hospital. The South-Transdanubian Region (STR, consisting of three counties, 22 small regions, 444 zipcode areas and 643 settlements) with 1 million inhabitants was the study area. All the studied parameters had significant spatial variability at all levels of aggregation. Beyond the relatively low average mortality risk in the STR, mortality clusters and increasing time trend were identified in certain areas. The mortality and the usage of mammography were inversely correlated with the indices of deprivation. These factors explain 64.5 and 17.5% of the whole variability of local mortality risks at the level of settlements and small regions. The explanatory role of these factors was similarly high for usage of mammography as well (40.2 and 52.6% for small regions and zipcode areas). The factors having the strongest influence were the population size (in settlement level mortality model), ratio of gypsies (in small region level mortality and mammography usage models) and ratio of Germans (in mammography usage model for zipcode areas). Inserting the counties' approaches for screening organisation into the model, it revealed that the population based screening organisation applied in Tolna county has the highest influence being 4.4 times stronger than the most important socio-economic factors.Altogether,it seems that the monitoring of spatial inequalities could improve the performance of breast cancer control identifying the populations with special needs, and there is a need to explore the pathways by which the socio-economic factors can exert their profound influence on the epidemiological status. Moreover, since the results clearly demonstrated that it is possible to achieve relatively high screening participation rates in Hungarian economical and legislative circumstances, the application of this successful method should be encouraged in other areas with low performance screening system.     

Synthesis,characterization, in silico ADMET prediction,and protein binding analysis of a novel zinc(II) Schiff-base complex: Application of multi-spectroscopic and computational techniques

By reaction of 1,2-diaminocyclohexane with the 2,3-butanedione monoxime in the presence of ZnCl₂, a new Schiff base complex was obtained. This complex was characterized by elemental analyses, FT-IR, ¹H NMR, UV–Vis, and conductivity measurements. The reactivity of this complex to human serum albumin (HSA) under simulative physiological conditions was studied by spectroscopic and molecular docking analysis. Experimental results at various temperatures indicated that the intrinsic fluorescence of protein was quenched through a static quenching mechanism. The negative value of enthalpy change and positive value of entropy change indicated that both hydrogen bonding and hydrophobic forces played a major role in the binding of Zn(II) complex to HSA. FT-IR, three-dimensional fluorescence, and UV–Vis absorption results showed that the secondary structure of HSA changed after Zn(II) complex bound to protein. The binding distance was calculated to be 4.96 nm, according to fluorescence resonance energy transfer. Molecular docking results confirmed the spectroscopic results and showed that above complex is embedded into subdomain IIA of protein. All these experimental and computational results clarified that Zn(II) complex could bind with HSA effectively, which could be a useful guideline for efficient Schiff-base drug design.     

Palladium(II) complexes of biorelevant ligands. Synthesis,structures, cytotoxicity and rich DNA/HSA interaction studies

In this work, a pair of new palladium(II) complexes, [Pd(Gly)(Phe)] and [Pd(Gly)(Tyr)], (where Gly is glycine, Phe is phenylalanine, and Tyr is tyrosine) were synthesized and characterized by UV–Vis, FT-IR, elemental analysis, ¹H-NMR, and conductivity measurements. The detailed ¹H NMR and infrared spectral studies of these Pd(II) complexes ascertain the mode of binding of amino acids to palladium through nitrogen of -NH₂ and oxygen of -COO^? groups as bidentate chelates. The Pd(II) complexes have been tested for in vitro cytotoxicity activities against cancer cell line of K₅₆₂. Interactions of these Pd(II) complexes with CT-DNA and human serum albumin were identified through absorption/emission titrations and gel electrophoresis which indicated significant binding proficiency. The binding distance (r) between these synthesized complexes and HSA based on Forster?s theory of non-radiation energy transfer were calculated. Alterations of HSA secondary structure induced by complexes were confirmed by FT-IR measurements. The results of emission quenching at three temperatures have revealed that the quenching mechanism of these Pd(II) complexes with CT-DNA and HSA were the static and dynamic quenching mechanism, respectively. Binding constants (K_b), binding site number (n), and the corresponding thermodynamic parameters were calculated and revealed that the hydrogen binding and hydrophobic forces played a major role when Pd(II) complexes interacted with DNA and HSA, respectively. We bid that [Pd(Gly)(Phe)] and [Pd(Gly)(Tyr)] complexes exhibit the groove binding with CT-DNA and interact with the main binding pocket of HSA. The complexes follow the binding affinity order of [Pd(Gly)(Tyr)] > [Pd(Gly)(Phe)] with CT-DNA- and HSA-binding.     

Fine-tuning the physicochemical properties of peptide-based blood–brain barrier shuttles

N-methylation is a powerful method to modify the physicochemical properties of peptides. We previously found that a fully N-methylated tetrapeptide, Ac-(N-MePhe)₄-CONH₂, was more lipophilic than its non-methylated analog Ac-(Phe)₄-CONH₂. In addition, the former crossed artificial and cell membranes while the latter did not. Here we sought to optimize the physicochemical properties of peptides and address how the number and position of N-methylated amino acids affect these properties. To this end, 15 analogs of Ac-(Phe)₄-CONH₂ were designed and synthesized in solid-phase. The solubility of the peptides in water and their lipophilicity, as measured by ultra performance liquid chromatography (UPLC) retention times, were determined. To study the permeability of the peptides, the Parallel Artificial Membrane Permeability Assay (PAMPA) was used as an in vitro model of the blood–brain barrier (BBB). Contrary to the parent peptide, the 15 analogs crossed the artificial membrane, thereby showing that N-methylation improved permeability. We also found that N-methylation enhanced lipophilicity but decreased the water solubility of peptides. Our results showed that both the number and position of N-methylated residues are important factors governing the physicochemical properties of peptides. There was no correlation between the number of N-methylated amide bonds and any of the properties measured. However, for the peptides consecutively N-methylated from the N-terminus to the C-terminus (p1, p5, p11, p12 and p16), lipophilicity correlated well with the number of N-methylated amide bonds and the permeability of the peptides. Moreover, the peptides were non-toxic to HEK293T cells, as determined by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay.