Binding site and interlobe interactions of the ionotropic glutamate receptor GluK3 ligand binding domain revealed by high resolution crystal structure in complex with (S)-glutamate

Ionotropic glutamate receptors (iGluRs) are involved in excitatory signal transmission throughout the central nervous system and their malfunction is associated with various health disorders. GluK3 is a subunit of iGluRs, belonging to the subfamily of kainate receptors (GluK1–5). Several crystal structures of GluK1 and GluK2 ligand binding domains have been determined in complex with agonists and antagonists. However, little is known about the molecular mechanisms underlying GluK3 ligand binding properties and no compounds displaying reasonable selectivity towards GluK3 are available today. Here, we present the first X-ray crystal structure of the ligand binding domain of GluK3 in complex with glutamate, determined to 1.6 Å resolution. The structure reveals a conserved glutamate binding mode, characteristic for iGluRs, and a water molecule network in the glutamate binding site similar to that seen in GluK1. In GluK3, a slightly lower degree of domain closure around glutamate is observed compared to most other kainate receptor structures with glutamate. The volume of the GluK3 glutamate binding cavity was found to be of intermediate size between those of GluK1 and GluK2. The residues in GluK3 contributing to the subfamily differences in the binding sites are primarily: Thr520, Ala691, Asn722, Leu736 and Thr742. The GluK3 ligand binding domain seems to be less stabilized through interlobe interactions than GluK1 and this may contribute to the faster desensitization kinetics of GluK3.     

New data on osteostracans (Agnatha) from the Lower Devonian of the Severnaya Zemlya Archipelago

The osteostracans Reticulaspis menneri gen. et sp. nov. and Nucleaspis unica gen. et sp. nov. from the Lower Devonian Severnaya Zemlya Formation of October Revolution Island of the Severnaya Zemlya Archipelago are described. The perfect preservation of the exoskeleton structure of Reticulaspis menneri allows the design and growth mode of the exoskeleton, with a continuous reticular dental surface of the cephalothoracic shield to be characterized. Nucleaspis unica is represented by a juvenile stage, which is extremely scarce in this vertebrate group. The preoccupied generic name Ungulaspis Afanassieva et Karatajūt?-Talimaa, 1998 is replaced by Paraungulaspis.     

Effect of genotype and medium composition on linseed (<Emphasis Type="Italic">Linum usitatissimum</Emphasis>) ovary culture

Breeding linseed (Linum usitatissimum L.) using haploid techniques allows breeders to develop new cultivars in a shorter time period. Many research groups successfully created new linseed genotypes through anther culture; however ovary culture has been the subject of only a few earlier studies. In the present study, the effect of genotype and growth regulators combination on callus induction and shoots regeneration in ovary culture of nine commercially important linseed cultivars was investigated. Ovaries were cultured on modified MS medium supplemented with three different combinations of plant growth regulators. Variable callogenic responses were expressed by all of the genotypes tested on different induction media. The results suggested that specific combination of growth regulators for callus induction must be designed for each genotype. Shoot regeneration from ovary derived callus is a critical phase of the whole gynogenetic process. Differences in adventitious shoot formation frequency among genotypes were demonstrated and four responsive genotypes have been selected. Ovary derived callus from cultivar ‘Mikael’ manifested the highest adventitious shoot formation frequency with a high number of shoots per explant. The optimum ratio of growth regulators for shoot regeneration was shown to depend on the genotype. Cultivars ‘Linola’, ‘Mikael’ and ‘Szaphir’ showed the highest shoot regeneration frequency when callus had originated on induction medium supplemented with 2 mg L⁻¹ BAP and 2 mg L⁻¹ NAA, while combination of 1 mg L⁻¹ BAP and 2 mg L⁻¹ IAA promoted shoot formation in ovary-derived callus of ‘Barbara’. The highest rate of shoots per explant has been obtained in second subculture.     

Redox reactions of the FAD-containing apoptosis-inducing factor (AIF) with quinoidal xenobiotics: a mechanistic study

Mitochondrial apoptosis-inducing factor (AIF) is a FAD-containing protein that under certain conditions translocates to the nucleus and causes a programmed cell death, apoptosis. The apoptogenic action of AIF is redox controlled as the NADH-reduced AIF dimer has lower affinity for DNA than the oxidized monomer. To gain further insights into the mechanism of AIF, we investigated its interaction with a series of quinone oxidants, including a number of anticancer quinones. Our data indicate that the NADH:quinone oxidoreduction catalyzed by AIF follows a “ping-pong” scheme, with the reductive half-reaction being rate-limiting and the FADH⁻–NAD⁺ charge-transfer complex serving as an electron donor. AIF is equally reactive toward benzo- and naphthoquinones, but may discriminate structures with a higher number of aromatic rings. The reactivity of quinones is mainly defined by their one-electron reduction potential, whereas the size and nature of the substituents play a minor role. AIF is unlikely to significantly contribute to bioreductive activation of low-potential quinoidal anticancer quinones. However, high-potential quinones, e.g. a toxic natural compound naphthazarin, maintain AIF in the oxidized state when a significant excess of NADH is present. Thus, these compounds may prevent the accumulation of the reduced form of AIF in vivo, and enhance AIF-mediated apoptosis.     

LED irradiance level affects growth and nutritional quality of Brassica microgreens

This study examines the effect of irradiance level produced by solid-state light-emitting diodes (LEDs) on the growth, nutritional quality and antioxidant properties of Brassicaceae family microgreens. Kohlrabi (Brassica oleracea var. gongylodes, ‘Delicacy Purple’) mustard (Brassica juncea L., ‘Red Lion’), red pak choi (Brassica rapa var. chinensis, ‘Rubi F₁’) and tatsoi (Brassica rapa var. rosularis) were grown using peat substrate in controlled-environment chambers until harvest time (10 days, 21/17°C, 16 h). A system of five lighting modules with 455, 638, 665 and 731 nm LEDs at a total photosynthetic photon flux densities (PPFD) of 545, 440, 330, 220 and 110 µmol m^?2s^?1 respectively were used. Insufficient levels of photosynthetically active photon flux (110 µmol m^?2 s^?1) suppressed normal growth and diminished the nutritional value of the Brassica microgreens studied. In general, the most suitable conditions for growth and nutritional quality of the microgreens was 330–440 µmol m^?2 s^?1 irradiation, which resulted in a larger leaf surface area, lower content of nitrates and higher total anthocyanins, total phenols and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging capacity. High light levels (545 µmol m^?2 s^?1), which was expected to induce mild photostress, had no significant positive impact for most of investigated parameters.     

Fatty acid amide hydrolase is a key regulator of endocannabinoid-induced myocardial tissue injury

Previous studies have suggested that increased levels of endocannabinoids in various cardiovascular disorders (e.g., various forms of shock, cardiomyopathies, atherosclerosis) through the activation of CB(1) cannabinoid receptors may promote cardiovascular dysfunction and tissue injury. We have investigated the role of the main endocannabinoid anandamide-metabolizing enzyme (fatty acid amide hydrolase; FAAH) in myocardial injury induced by an important chemotherapeutic drug, doxorubicin (DOX; known for its cardiotoxicity mediated by increased reactive oxygen and nitrogen species generation), using well-established acute and chronic cardiomyopathy models in mice. The DOX-induced myocardial oxidative/nitrative stress (increased 4-hydroxynonenal, protein carbonyl, and nitrotyrosine levels and decreased glutathione content) correlated with multiple cell death markers, which were enhanced in FAAH knockout mice exhibiting significantly increased DOX-induced mortality and cardiac dysfunction compared to their wild type. The effects of DOX in FAAH knockouts were attenuated by CB(1) receptor antagonists. Furthermore, anandamide induced enhanced cell death in human cardiomyocytes pretreated with FAAH inhibitor and enhanced sensitivity to ROS generation in inflammatory cells of FAAH knockouts. These results suggest that in pathological conditions associated with acute oxidative/nitrative stress FAAH plays a key role in controlling the tissue injury that is, at least in part, mediated by the activation of CB(1) receptors by endocannabinoids.     

The use of benthic macroinvertebrate families for river quality assessment in Lithuania

The aim of this study was to evaluate the influence of environmental factors on the distribution of macroinvertebrate taxa in different sized lowland Lithuanian rivers. A secondary aim was to assess ecological river quality and to determine the most suitable biotic index. A final aim was to determine the most appropriate macroinvertebrate families for river quality assessment in Lithuania. Species composition and quantitative characteristics of benthic macroinvertebrate communities have been investigated using standard kick-sampling method by a standard hand net in 24 different river sites in spring. Physical and chemical environmental parameters were measured in the same study site as the macroinvertebrate sampling. A total of 186 taxa representing 66 families or higher taxonomic ranks of benthic macroinvertebrates have been identified. Water temperature and current velocity influenced the highest number of ivestigated families. Seven of the most tolerant and eleven of the most sensitive macroinvertebrate taxa for hydrochemical parameters related with organic pollution were determined. The DSFI method was founded to be the best index for assessment of ecological status for Lithuanian rivers until more accurate estimation method will be created.     

<Emphasis Type="Italic">Aeromonas salmonicida</Emphasis> infected fish transfer disease to healthy fish <Emphasis Type="Italic">via</Emphasis> water

Experimental studies of infection transmission via water from infected to healthy fish were conducted. The dark-brown bacterial colonies typical for Aeromonas salmonicida on tryptone soya agar (TSA) have been isolated and counted (from 3.0±0.6×10² to 3.5±0.5×10⁵ c.f.u. g⁻¹) from the internal organs of naturally infected (NI) and experimentally infected (EI) perch and sea trout. No significant differences in dark-brown bacterial counts were detected between EI perch and EI sea trout. The assessment and comparison of the alterations of the biological parameters of EI European perch and sea trout with bacterium Aeromonas salmonicida subsp. salmonicida with naturally infected perch were conducted. No mortality was recorded in groups of EI perch and sea trout. Whereas, the mortality of NI perch (collected from the main sites of outbreak of disease) was observed from the second day of the experiments. Changes in morphophysiological parameters of EI perch and sea trout were similar. Different alterations in blood cell parameters of EI fish were observed, and the most noticeable was the decrease (P≤0.01) in white blood cell count (WBC) of EI perch and sea trout. Based on these results it can be deduced that there is infection transmission of bacterium A. salmonicida from European perch via water to other fish species.     

Relationships of Oxygen Consumption, Hemodynamics, and Heart Rate Variability with the Functional Class of Coronary Artery Disease Patients

The functional classes (FCs) established according to the criteria of the New York Heart Association were tested for association with oxygen consumption, the state of central hemodynamics, and heart rate (HR) variability in coronary artery disease (CAD) patients. Oxygen consumption, central hemodynamics, and HR variability at rest and during exercise were assayed in 146 CAD patients and 30 healthy individuals (the control group). It was established that the peak oxygen consumption (VO_2max), anaerobic threshold, pulmonary ventilation, systolic and minute blood volume at the threshold load (TL), and HR variability in a supine position significantly decrease and the total vascular peripheral resistance at rest and during exercise increases with increasing FC in CAD patients. The closest correlation of FC was revealed with physical working capacity, anaerobic threshold, age, and peak oxygen consumption. Moderate correlations were established with the chronotropic function of the heart at the threshold load, HR variability, the high- and low-frequency components of the cardiac rhythm at TL, pulmonary ventilation, stroke volume at rest and at TL, and the carbon dioxide ventilation equivalent at TL. In healthy individuals, the peak oxygen consumption closely correlated with the HR variability, the very low frequency component at TL, and physical capacity. With an increase in FC in CAD patients, peak oxygen consumption became more tightly associated with the chronotropic function and the hemodynamic components at TL than with the HR variability in a supine position or at TL.     

<Emphasis Type="Italic">In vitro</Emphasis> culture of <Emphasis Type="Italic">Chrysanthemum</Emphasis> plantlets using light-emitting diodes

Effects of illumination spectrum on the morphogenesis of chrysanthemum plantlets (Chrysanthemum morifolium Ramat. ‘Ellen’) grown in vitro were studied using an illumination system consisting of four groups of light-emitting diodes (LEDs) in the following spectral regions: blue (450nm), red (640nm), red (660nm), and far-red (735nm). Taking into account all differences in shoot height, root length, and fresh and dry weight (FW and DW, respectively), observed while changing the total photon flux density (PFD), the optimal total PFD for growth of chrysanthemum plantlets in vitro was estimated. For 16 h photoperiod and typical fractions of the spectral components (14%, 50%, 28%, and 8%, respectively), the optimal total PFD was found to be 40 μmol m⁻² s⁻¹. Our study shows that the blue component in the illumination spectrum inhibits the plantlet extension and formation of roots and simultaneously increases the DW to FW ratio and content of photosynthetic pigments. We demonstrate photomorphogenetic effects in the blue region and its interaction with the fractional PFD of the far-red spectral component. Under constant fractional PFD of the blue component, the root number, length of roots and stems, and fresh weight of the plantlets have a correlated nonmonotonous dependence on the fractional PFD of the far-red component.