Structural and Functional State of Thylakoids in a Halophyte Suaeda altissima before and after Disturbance of Salt–Water Balance by Extremely High Concentrations of NaCl

Halophyte plants Suaeda altissima L. were grown in water culture at different concentrations of NaCl in the medium, and their leaves were sampled to examine the ultrastructure of chloroplasts. In parallel tests, the functional state of chloroplasts was assessed from parameters of chlorophyll fluorescence. In addition the effects of NaCl on plant growth and on the contents of Na⁺, K⁺, and water in organs of S. altissima were investigated. At a wide range of external salt concentrations (0–750 mM NaCl), S. altissima plants retained the chloroplast ultrastructure and photosynthetic function in an intact condition. The impairment of thylakoid ultrastructure and the accompanying increase in nonphotochemical quenching of excited states of chlorophyll was only observed at an extremely high concentration of NaCl in the medium (1 M) that led to disruption of ionic homeostasis and lowered water content in tissues.     

High Yield Elution of Proteins from Sodium Dodecyl Sulfate–Polyacrylamide Gels at the Low-Picomole Level. Application to N-Terminal Sequencing of a Scarce Protein and to In-Solution Biological Activity Analysis of On-Gel Renatured Proteins

A simple, reliable procedure for practically quantitative (90–98%) and fast (<30 min) elution of proteins from SDS-PA gels is described with reproducible recoveries in the range from 100 to 1 pmol per band, which does not require the inclusion of detergents in the elution buffer. It consists in the combination of (1) highly sensitive on-gel protein detection (50 mol per band) with imidazole-SDS-zinc (reverse staining), (2) crushing of the protein band to produce 32-m gel particles, and (3) vortexing of the slurry in a solution of a zinc-complexing agent, e.g. glycine 0.5 M or EDTA 100 mM (100 l for a 100-pmol BSA band), at room temperature. Eluted proteins can be directly analyzed by RP-HPLC, quantitatively loaded onto a PVDF membrane, or, provided that they are previously renatured on-gel, analyzed by biological activity tests. The application of the procedure to in-solution enrichment of scarce proteins for N-terminal analysis is shown.     

Simulations of the Ion Current to a Probe in Plasma with Allowance for Ionization and Ion–Neutral Collisions: II. Cylindrical Probe

The ion current to a cylindrical probe is considered with allowance for volume ionization, ion–neutral collisions, and the ion orbital moment. A model based on the molecular dynamics method and applicable in a wide range of plasma parameters (r_p/λ_D= 0.01–100, r_i/λ_D= 0.002–200, ν_i/ω_0i= 0.01–0.05, and T_i/T_e = 0?0.01) is proposed A convenient representation of the dependence of the relative ion current density on the Langmuir coefficient β² and a technique for determining the plasma density from simulation results are offered.     

Evidence that binding to the S2-subsite of papain may be coupled with catalytically relevant structural change involving the cysteine-25–histidine-159 diad. Kinetics of the reaction of papain with a two-protonic-state reactivity probe containing a hydrophobic side chain

A method is proposed by which site-specific reactivity probes that exhibit different reactivities in two ionization states can be used to detect association-activation phenomena that involve repositioning of acid/base groups in enzyme active centres. The pH-dependences of the apparent second-order rate constants (k) for the reactions of the thiol group of papain (EC 3.4.22.2) with a series of two-protonic-state reactivity probes are compared. The short-chain probes, 2,2'-dipyridyl disulphide and n-propyl 2-pyridyl disulphide, react at pH6 in adsorptive complexes and/or transition states with geometries that do not permit hydrogen-bonding of the pyridyl nitrogen atom with the active-centre imidazolium ion, as evidenced by the rate minima at pH6 and the rate maxima at pH4 provided by reagent protonation. Only when the probe molecule, e.g. 4-(N-aminoethyl 2'-pyridyl disulphide)-7-nitrobenzo-2-oxa-1,3-diazole [compound(III)], contains a long hydrophobic side chain is the reaction characterized by maximal rates at about pH6, as in the acylation step of the catalytic act (at pH6, k(compound III)/k(2,2'-dipyridyl disulphide) approximately 100). It is proposed that this striking difference in profile shape may result from binding of the hydrophobic side chain of compound (III) possibly in the S(2)-subsite of papain, which promotes a change in catalytic-site geometry involving repositioning of the imidazolium ion of histidine-159 and hydrogen-bonding with the N atom of the leaving group, as has been postulated to occur in the acylation step of substate hydrolysis.     

Phyto- and Zooplankton of Open Overgrown Shallows of the Rybinsk Reservoir Adjacent to a Mixed Grey Heron–Great Egret (<Emphasis Type="Italic">Ardea cinerea</Emphasis> L. and <Emphasis Type="Italic">A. alba</Emphasis> L.) Colony at a High Water Level

Phyto- and zooplankton communities developing in the open overgrown shallows of the Rybinsk Reservoir under the influence of a bird colony are shown to lose specific traits of quantitative characteristics and structure fluctuation in response to the elevated water level in contrast to previous findings. Possible reasons, such as the decomposition of sunken coastal vegetation playing the main role, accompanied by the development of semiaquatic macrophytes and trophic relations between algae and invertebrates, are discussed. It is found that increased atmospheric precipitation leads to an increased abundance of Copepoda in the zooplankton. Presumably, this effect correlates with more rapid and intensive inflow of avian vitality products rich in nitrogen into the water, resulting in a change in the stoichiometric ratio of nitrogen and phosphorus in zooplankton food objects to values favorable for copepods.