Active papain renatured and processed from insoluble recombinant propapain expressed in Escherichia coli.

For the first time the pro-form of a recombinant cysteine proteinase has been expressed at a high level in Escherichia coli. This inactive precursor can subsequently be processed to yield active enzyme. Sufficient protein can be produced using this system for X-ray crystallographic structure studies of engineered proteinases. A cDNA clone encoding propapain, a precursor of the papaya proteinase, papain, was expressed in E. coli using a T7 polymerase expression system. Insoluble recombinant protein was solubilized in 6 M guanidine hydrochloride and 10 mM dithiothreitol, at pH 8.6. A protein-glutathione mixed disulphide was formed by dilution into oxidized glutathione and 6 M GuHCl, also at pH 8.6. Final refolding and disulphide bond formation was induced by dilution into 3 mM cysteine at pH 8.6. Renatured propapain was processed to active papain at pH 4.0 in the presence of excess cysteine. Final processing could be inhibited by the specific cysteine proteinase inhibitors E64 and leupeptin, but not by pepstatin, PMSF or EDTA. This indicates that final processing was due to a cysteine proteinase and suggests that an autocatalytic event is required for papain maturation.     

Interrelations between sympathetic adrenal and opioid systems--regulatory mechanism determining cardiac resistance to stress damage]

The present paper has analyzed relationship between sympatico-adrenal and opioid systems in the pathogenesis of stress heart damage. Based on the our own results and other investigator data the authors make a conclusion that namely relationship between opioid and sympatico-adrenal systems both on the level of the brain and on the periphery determines a degree of the heart resistance to the injury action of severe stress. Myocardial protection by opioids at stress was found to be mediated by the peripheral mu-opioid receptors and was associated with decrease in an activity of sympatico-adrenal system and a inhibition of its effector part. On contrary central opioid system activation leads to an increase in stress heart damage via an increase in sympathetical influence on the myocardium.     

Biosorption of Cd, Zn and Cu by Sargassum sp. waste biomass

Biosorption is becoming an important component in the integrated approach to the treatment of aqueous effluents. The economics of biomass technical applications are improved by using waste biomass instead of purposely-produced biomass. Biomass derived from an alginate extraction industry - Sargassum sp. - was examined for its ability to function as a biosorbent for metals such as cadmium, zinc and copper. For use in column applications, biomass should be immobilized. To the algae reinforcement, the biomass was embedded in polyethleneimine (PEI), followed by glutaraldehyde crosslinking. Equilibrium Zn and Cu isotherms were analysed using the immobilization ratio that showed the best Cd performance. Either Freundlich or Langmuir models can describe the passive biosorption equilibrium of cadmium, zinc and cooper. The preference for this series of metals by the biomass was found to be Cd > Zn > Cu, with maximum uptake values of 157.8, 118.5 and 77.4 mg/g dry weight biomass for Cd, Zn and Cu. respectively. The metal binding capacity by non-living biomass is an important quality for industrial use.     

The lagena (the third otolith endorgan in vertebrates)

In this review, the structure and functions of the lagena (the third otolith organ) in an evolutionary lineage of the vertebrates are described and discussed. The lagenar macula appears first in the posterior part of the sacculus of elasmobranchs; in these animals, the lagena is considered to be involved in the balance support (orientation with respect to the gravitation force). The lagena as a separate endorgan has been described in teleost fishes; in some species, the lagena is connected with the sacculus, while in other species the interrelations of these structures can be dissimilar. The lagena supplements the functions of the sacculus; in fishes (animals with no special organ of hearing), it is involved in discrimination of sound oscillations, identification of the gravitation vector, and orientation in the course of movements within the vertical plane. In amphibians, the lagena is localized in the posterior part of the sacculus, near the auditory structures; it performs mostly vestibular and (to a much lesser extent) auditory functions. In amniotes, the lagena was first separated from the sacculus; it is localized in the cochlear canal, distally with respect to the hearing organ. Information on the functions of the lagena in amniotes is rather limited and contradictory. Central projections of this organ have been examined practically only in birds. Lagenar afferents project to the vestibular nuclei and cerebellum, while some fibers come to the auditory nuclei of the medulla. The lagena in birds can be related to their navigation abilities (birds are supposed to be capable of orienting within the magnetic field of the Earth due to the magnetic properties of the lagenar otoconia; this structure can also provide detection of movements along the vertical axis. The close proximity between the otolithic and auditory endorgans in the cochlear canal of amniotes can be indicative of the functional significance of these interrelations. This aspect, however, remains at present undiscovered. In mammals (except Monotremata), there is no lagena as an independent endorgan. Neirofiziologiya/Neurophysiology, Vol. 40, No. 2, pp. 160–178, March–April, 2008.     

Behavior of a dust grain in the double layer of an electric probe in a gas-discharge plasma

Equations for the motion of an individual dust grain in the double layer of a negatively charged cylindrical probe in a glow discharge plasma are derived and solved numerically. The distribution of the electric potential near the probe is determined, and the grain charge is calculated as a function of the distance from the probe for different probe potentials. The trajectories of grains with different initial energies are traced. An analysis of the grain trajectories shows that, at a certain distance from the probe, high-energy grains may be recharged; i.e., the grain charge may change sign. The grains are found to have no direct effect on the probe current in a dusty plasma of a glow discharge.     

EEG spatial organization during intense hyperventilation (cyclic breathing): II. EEG correlates of psychovisceral phenomena

EEG correlates of two main effects of long-term hyperventilation (cyclic breathing similar to rebirthing), namely, psychotic manifestations and sensations in internal organs, are considered. Standard EEGs of 44 subjects were recorded and subsequently treated by multiparametric methods. It was shown that, in the case of psychotic phenomena (hallucinations, visual and auditory images, different scenes, virtual travels, out-of-body experiences, etc.), the changes in the spatial organization of bioelectric potentials as compared with the baseline have different patterns than in the case of sensations in internal organs. The changes are observed in the frontal regions of the cortex: a decrease in spatial synchronization (linear processes) during an increase in spatial disarray (nonlinear processes) of bioelectric potentials and the intensifying of high-frequency β activity. If sensations in internal organs appear, a decrease in the above parameters was observed in the right hemisphere and in posterior cortical areas. The data are discussed from the viewpoint of qualitative specificity of changes in functional states during an altered state of consciousness with manifestations of different psychovisceral phenomena.     

Rare earth thiocyanate complexes with tripiperidinophosphine oxide: synthesis and characterization. Crystal structure of Pr(SCN)3(tpppO)3

The synthesis and characterization of rare-earth (La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu and Y) thiocyanate adducts with tripiperidinophosphine oxide (tpppO) with general formula (RE)(SCN)₃(tpppO)₃ are reported. Conductance measurements in acetonitrile indicate the non-electrolytic nature of the complexes. Infrared absorption spectra evidence that the SCN⁻ ion coordinates through the nitrogen atom (isothiocyanate form) and that tpppO coordinates through the phosphoryl oxygen. X-ray powder patterns suggest the existence of three different crystal forms: (1) La; (2) an isomorphous series including Ce, Nd and Pr; and (3) another isomorphous series, including Sm, Gd, Eu, Ho, Er, Tb, Lu and Y. The visible spectra of the Nd adduct and the calculated parameters β = 0.98, b^1/2 = 0.072 and δ = 1.06 indicate that the metal-ligand bonds are essentially electrostactic. The emission spectra of the Eu compound showed ⁵D₀ → ⁷F_J bands (J = 0, 1, 2), suggesting a C_3v symmetry for the coordination polyhedron. The lifetime of the ⁵D₀ state is 1.28 ms. The emission spectra of the Tb complex presented ⁵D₄ → ⁷F_J bands (J = 4, 5, 6) and the Dy complex showed the ⁴F_9/2 → ⁶H_13/2 band. The structure of the Pr complex showed that the coordination polyhedron is a trigonal antiprism, with the isothiocyanate anions in one base and three tpppO ligands in the other. Thermal analyses (TG-DTG) were carried out for the Ce, Nd and Gd adducts. Mass losses start between 250 and 334 °C. The final residues at 1300 °C are the corresponding phosphates.     

Long term effects of Chernobyl contamination on DNA repair function and plant resistance to different biotic and abiotic stress factors

Thirty years after the Chernobyl explosion we still lack information regarding the genetic effects of radionuclide contamination on the plant population. For example, are plants adapting to the low dose of chronic ionising irradiation and showing improved resistance to radiation damage? Are they coping with changing/increased pathogenicity of fungi and viruses in the Chernobyl exclusion (ChE) zone? Are plant populations rapidly accumulating mutational load and should we expect rapid micro-evolutionary changes in plants in the Chernobyl area? This review will try to summarise the current knowledge on these aspects of plant genetics and ecology and draw conclusions on the importance of further studies in the area around Chernobyl.     

Effect of Cimetidine and other antihistaminics on the elimination of aminopyrine,phenacetin and caffein

Cimetidine is widely prescribed for the treatment of peptic ulcer disease and has recently been shown to inhibit the metabolism of warfarin, antipyrine and diazepam. To further examine this phenomenon we investigated the effect of various doses of cimetidine and other related drugs on ¹⁴C-aminopyrine, ¹⁴C-phenacetin and ¹⁴C-caffeine breath tests. Cimetidine caused a dose-related inhibition of the metabolism of aminopyrine and caffeine but had no effect on the phenacetin breath test. Metiamide, H₁-antihistamines, phenothiazines and local anesthetics also inhibited the aminopyrine breath test. Cyproheptadine had no effect on either phenacetin or caffeine elimination. This study demonstrates a complex drug-drug interaction which may have widespread clinical implications.