Inhibitory Modulation by Sodium Ions of the N-Methyl-D-Aspartate Recognition Site in Brain Synaptic Membranes

Specific binding of radiolabeled L-glutamic acid (Glu) was examined using rat brain synaptic membranes treated with a low concentration of Triton X-100. The binding drastically increased in proportion to increasing concentrations of the detergent used up to 0.1%. Addition of 100 mM sodium acetate significantly potentiated the binding in membranes not treated with Triton X-100, whereas it markedly inhibited the binding in Triton-treated membranes. The binding in Triton-treated membranes was inversely dependent on incubation temperature and reached a plateau within 10 min after the initiation of incubation at 2 degrees C, whereas the time required to attain equilibrium at 30 degrees C was less than 1 min. Sodium acetate invariably inhibited the binding detected at both temperatures independently of the incubation time via decreasing the affinity for the ligand. The binding was significantly displaced by agonists and antagonists for an N-methyl-D-aspartate (NMDA)-sensitive subclass of brain excitatory amino acid receptors, but not by those for the other subclasses. Inclusion of sodium acetate reduced the potencies of NMDA agonists to displace the binding without virtually affecting those of NMDA antagonists. Moreover, sodium ions inhibited the ability of Glu to potentiate the binding of N-[3H] [1-(2-thienyl)cyclohexyl]piperidine to open NMDA channels in Triton-treated membranes. These results suggest that sodium ions may play an additional modulatory role in the termination process of neurotransmission mediated by excitatory amino acids via facilitating a transformation of the NMDA recognition site from a state with high affinity for agonists to a state with low affinity.     

Ca2+-Independent, Ca2+-Dependent, and Carbachol- Mediated Arachidonic Acid Release from Rat Brain Cortex Membrane

Synaptoneurosomes obtained from the cortex of rat brain prelabeled with [14C]arachidonic acid [( 14C]AA) were used as a source of substrate and enzyme in studies on the regulation of AA release. A significant amount of AA is liberated in the presence of 2 mM EGTA, independently of Ca2+, primarily from phosphatidic acid and polyphosphoinositides (poly-PI). Quinacrine, an inhibitor of phospholipase A2 (PLA2), suppressed AA release by about 60% and neomycin, a putative inhibitor of phospholipase C (PLC), reduced AA release by about 30%. An additive effect was exhibited when both inhibitors were given together. Ca2+ activated AA release. The level of Ca2+ present in the synaptoneurosomal preparation (endogenous level) and 5 microM CaCl2 enhance AA liberation by approximately 25%, whereas 2 mM CaCl2 resulted in a 50% increase in AA release relative to EGTA. The source for Ca(2+)-dependent AA release is predominantly phosphatidylinositol (PI); however, a small pool may also be liberated from neutral lipids. Carbachol, an agonist of the cholinergic receptor, stimulated Ca(2+)-dependent AA release by about 17%. Bradykinin enhanced the effect of carbachol by about 10-15%. This agonist-mediated AA release occurs specifically from phosphoinositides (PI + poly-PI). Quinacrine almost completely suppresses calcium-and carbachol-mediated AA release. Neomycin inhibits this process by about 30% and totally suppresses the effect of bradykinin. Our results indicate that both phospholipases PLA2 and PLC with subsequent action of DAG lipase are responsible for Ca(2+)-independent AA release. Ca(2+)-dependent and carbachol-mediated AA liberation occurs mainly as the result of PLA2 action. A small pool of AA is probably also released by PLC, which seems to be exclusively responsible for the effect of bradykinin.     

Negative air ions as a source of superoxide

The physico-chemical characteristics and possible formation mechanisms of negative air ions are considered. It was found that the products of oxygen and nitrogen negative ionization reduce ferricytochromec and nitroblue tetrazolium, and that these reactions were inhibited by superoxide dismutase. The interaction of negatively ionized oxygen with water led to hydrogen peroxide accumulation, which was inhibited by tetranitromethane or catalase. Nitrogen ionization under these conditions caused the formation of the hydrated electron e _aq ^— and the superoxide anion O ₂ ^— . The data obtained indicate that the biological activity of negative air ions may be dependent on superoxide. The generation of reactive oxygen ions in the gas phase and also at a gas/water interface is described. A scheme for superoxide production under oxygen and nitrogen ionization is proposed.     

Glucose oxidase immobilized electrode for potentiometric estimation of glucose

Glucose oxidase has been immobilized onto a thin platinum strip, by co-crosslinking with bovine serum albumin and glutaraldehyde. The retention of redox characteristics of glucose oxidase has been verified by cyclic voltammetry. The activity of the immobilized enzyme reduces to a quarter of its value when the enzyme is in solution but improves when coimmobilized with 1 urea. The potentiometric response builds up and remains stable after 100 s. It is sensitive to the thickness of the immobilizing matrix, pH and temperature. An improvement in the performance of the electrode has been achieved by coimmobilizing 2 urea and metal ions such as Mg²⁺ and Mn²⁺. The presence of Cu has been proved to be detrimental. The electrode has been calibrated in the 0.1–5.0 mM glucose concentration range. It gives a stable response for more than 50 independent assays and can be stored for 60 days without significant loss of function.     

Nutrient content and ionic pattern in beech (Fagus sylvatica L.) from natural stands in Eastern Austria and ecological implications

Macro and Micro cation content, concentration of water soluble cations and anions, as well as nitrogen content in beech leaves from a variety of forest stands in the eastern part of Austria have been determined. Special attention has been focused on the highly air polluted Vienna Woods. A broad natural variability in total content of minerals, as well as in nitrogen content could be observed. In general, cation nourishment has been found to be adequate on all stands. The pattern of water soluble i.e. physiologically active cations, especially with regard to divalent cations, varies broadly among beeches from different provenances: Mg/Mn ratios differ within three orders of magnitude between dolomite and highly acidic, nutrient poor stands over silicate rock. The term mineral deficiency limit should, therefore, be used very carefully with respect to single nutrients. Organic anions (quinate, malate, citrate) clearly dominate over inorganic anions. The relative amount of the latter increased from NO₃, Cl, inorganic phosphate to SO₄. A concentration of SO₄ is markedly in leaves within the direct reach of SO₂ sources, its level, however, remaining well below the organic anions. Slight evidence of an antagonistic behavior between SO₄ and organic acids could be found. Total nitrogen as well as the ratio insoluble:soluble nitrogen were tendencially higher in Vienna Wood beeches, indicating an additional use of the airborne nitrogen. The high flexibility of beech with respect to mineral content and ionic balance is discussed with regard to the ecological distribution of beech and the susceptibility to air pollution.     

Dependence of the photophobic response of the blue-green alga,Phormidium uncinatum,on cations

It is suggested that photophobic responses caused by a sudden step-down in light intensity require the presence of cations in the blue-green alga, Phormidium uncinatum.Drastic removal of cations abolishes the phobic response, which recovers after addition of Ca²⁺ ions. Calcium can be substituted for partially by other cations with an effectivity following the sequence Ca>Mg>Na>Ba>Co=0. During the photophobic response there is a 25% increase in ⁴⁵Ca binding by the cells related to a step-down in light intensity. Three seconds after a light-dark transition there is a sharp increase in the binding of labelled calcium, followed by a subsequent release.Flushing the filaments with high cation concentrations, esp. calcium causes a reversal of movement in the absence of a light stimulus similar to a photophobic reversal. This stimulus could trigger the same sequence of events in the transduction chain bypassing the primary photoresponse.Abbreviations EDTA Ethylene diaminetetraacetic acid - EGTA ethylene glycol-bis (2-aminoethylether) N,N tetraacetic acid     

Effect of cations on the structure of mung bean cytoplasmic ribosomes

The importance of the absolute and relative concentrations of monovalent and divalent cations and centrifugal speed (pressure) in the dissociation of mung bean 80S ribosomes has been examined. In the absence of Mg²⁺ ions, ribosome monomers yield 47S and 34S particles. Fixation with glutaraldehyde, however, indicates that this dissociation pattern is largely dependent upon high pressures developed during centrifugation and that in the absence of such artifacts the immediate product of Mg-free conditions is a 74S particle. Since 74S particles rapidly revert to the 80S form when Mg is replaced, this would appear to be a conformational change. Ribosomes were also dissociated in the presence of Mg²⁺ ions if the K⁺ ion concentration was raised. Three major particles were produced, 38S and 49S from the small ribosomal sub-unit and 60S from the large sub-unit. A proportion of the 80S monomer population is more resistant to dissociation. Experiments with puromycin indicate that the more resistant fraction probably represents ribosomes completed with nascent polypeptide resulting from polysome breakdown.     

Insight towards the conserved water mediated recognition of catalytic and structural Zn+2 ions in human Matrix Metalloproteinase-8 enzyme: A study by MD-simulation methods

Human matrix metalloproteinase-8 (hMMP-8) plays a important role in the progression of colorectal cancer, metastasis, multiple sclerosis and rheumetoid arthritis. Extensive MD-simulation of the PDB and solvated structures of hMMP-8 has revealed the presence of few conserved water molecules around the catalytic and structural zinc (ZnC and ZnS) ions. The coordination of two conserved water molecules (W and WS) to ZnS and the H-bonding interaction of WS to S151 have indicated the plausible involvement of that metal ion in the catalytic process. Beside this the coupling of ZnC and ZnS metal ions (ZnC – W_H (W₁)…..W₂ ….H₁₆₂ - ZnS) through two conserved hydrophilic centers (occupied by water molecules) may also provide some rational on the recognition of two zinc ions which were separated by ~13 Å in their X-ray structures. This unique recognition of both the Zn⁺² ions in the enzyme through conserved water molecules may be implemented/ exploited for the design of antiproteolytic agent using water mimic drug design protocol.     

Yeasts and yeast-like fungi in tap water and groundwater,and their transmission to household appliances

In the present study we describe the occurrence of fungi in 100 tap water and 16 groundwater samples from Slovenia. We used culture-dependent and culture-independent techniques. 28 fungal species belonging to 16 genera were isolated with selected culturing conditions, targeting human opportunistic yeasts and yeast-like fungi. Of special concern was the detection of Aureobasidium melanogenum, Exophiala dermatitidis, Rhinocladiella similis, Candida parapsilosis and Rhodotorula mucilaginosa. The DGGE analysis of ITS1 rDNA revealed from 6 to 16 bands hypothetically corresponding to different taxa, while pyrosequencing showed the presence of Aspergillus and Exophiala. According to the statistic machine learning methodology, the profile of fungi in water is determined by the concentration of calcium and magnesium ions and the presence of nitrate. Exophiala spp., C. parapsilosis and R. mucilaginosa are known as dominant contaminants of household appliances. It appears that they are transferred with water to dishwashers and washing machines, where they subsequently proliferate.     

Study of trivalent samarium ion embedded lithium‐based borate glass for high‐density optical memory devices

Sm³⁺ ions doped strontium lithium lead borate glasses (SLLB:Sm) were prepared using a conventional melt‐quenching technique. The glasses were analyzed using X‐ray diffractometry and Fourier transform infrared spectroscopy, optical absorption, fluorescence spectral analysis, and fluorescence lifetime decay. The Judd–Ofelt (J–O) parameters and radiative parameters of the SLLB:Sm10 glass (1.0 mol% Sm³⁺ ion‐doped glass) were calculated using J–O theory. From the emission spectra, among all the synthesized glass, SLLB:Sm10 glass had the highest emission intensity for ⁴G_5/2→⁶H_11/2 transition (610 nm). Emission parameters, such as stimulated emission cross‐section and optical gain bandwidth, were calculated. For all concentrations of Sm³⁺ ions, the decay profile showed an exponential nature and decreased when the Sm³⁺ ion concentration was increased due to a concentration quenching effect. This result suggests that the synthesized SLLB:Sm10 glass could be used for application in high‐density optical memory devices.