Stoichiometry of microsomal oxidation reactions. Distribution of redox-equivalents in monooxygenase and oxidase reactions catalyzed by cytochrome P-450

The stoichiometry of NADPH oxidation in rabbit liver microsomes was studied. It was shown that in uncoupled reactions cytochrome P-450, besides O2- generation catalyzes direct two- and four-electron reduction of O2 to produce H2O2 and water, respectively. With an increase in pH and ionic strength, the amount of O2 reduced via an one-electron route increases at the expense of the two-electron reaction. In parallel, with a rise in pH the steady-state concentration of the oxy-complex of cytochrome P-450 increases, while the synergism of NADPH and NADH action in the H2O2 formation reaction is replaced by competition. The four-electron reduction is markedly accelerated and becomes the main pathway of O2 reduction in the presence of a pseudo-substrate--perfluorohexane. Treatment of rabbit with phenobarbital, which induces the cytochrome P-450 isozyme specific to benzphetamine results in a 2-fold increase in the degree of coupling of NADPH and benzphetamine oxidation. The experimental results suggest that the ratio of reactions of one- and two-electron reduction of O2 is controlled by the ratio of rates of one- and two-electron reduction of cytochrome P-450. In the presence of pseudo-substrates cytochrome P-450 acts predominantly as a four-electron oxidase; one of possible reasons for the uncoupling of microsomal monooxygenase reactions is the multiplicity of cytochrome P-450 isozymes.     

Responses of receptive fields of frog retinal ganglion cells to the leading and trailing edges of moving stimuli

The position of on- and off-discharge centers in class 1 and 3 receptive fields of the frog retina was determined with the aid of moving bars of different lengths. On- and off-centers of receptive fields of the first group coincide, those of the second are spatially separate, and in fields of the 3rd group the discharge center of one contrast sign occupies the central position and discharge centers of the opposite sign are located at the periphery, to its right and left. Receptive fields of the frog retina thus have features which approximate them to the concentric receptive fields of geniculate neurons and the fields of the cat visual cortex. Asymmetry in the responses was found: during movement in opposite directions the distance between the discharge centers changed, during movement to one side only one of the peripheral centers was revealed, whereas during movement to the other side the second center was revealed on the opposite side of the receptive field. This asymmetry of spatiotemporal relations in the receptive fields is similar to that found in the fields of cortical neurons and is connected with their directional properties.Research Institute of Applied Mathematics and Cybernetics, N. I. Lobachevskii State University, Gor'kii. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 75–85, January–February, 1980.     

Restoration of morphological and functional integrity in the regenerating eye of the giant African land snail Achatina fulica

To determine whether vision returns to its original state following eye removal in Achatina fulica, light and electron microscope examinations, electrophysiological recordings and behavioural tests were carried out on the regenerating snails. Reparative morphogenesis can result in the restoration of the peripheral sense organ even in the absence of complete regrowth of the tentacle, but it can also lead to the formation of aberrant regenerates. We found that anatomically and ultrastructurally the eyes of the ‘most normal’ regenerates were basically the same as the original eyes. Under normal conditions each eye is composed of a principal and an accessory eye, both sharing a common cornea. The only difference between regenerated and native eyes is the smaller size of the former, as a result of a reduced number of retinal cells. Electroretinographic responses revealed that the molecular mechanism of phototransduction is restored, in principle, but that flicker fusion frequency in the regenerated eye is significantly lower than in the normal eye. The directional movement to a visual stimulus (a black stripe of 45° width) had not completely recovered even 6 months after amputation. This suggests that the central projections of the optic nerve had not become fully re‐established at the time of testing.     

Middle paleozoic waulsortian-type mud mounds in Southern Fergana (Southern Tien-Shan,commonwealth of independent states): The shallow-water atoll model

Summary Several Waulsortian-type mud mounds nearly 500 m thick and about 5 km long occur in the Middle Paleozoic carbonate section of the Aktur nappe in the mountains on the right bank of Isfara river. These buildups form a well developed barrier system that stretches along the South Ferganian carbonate platform margin and divides the carbonate complex into a fore-reef and a back-reef part. The time of the mounds' most active growth was from the Late Silurian (Ludlow) to the Middle Devonian (Eifel). Three main facies types can be recognized in the mud mounds: 1. micritic core facies, 2. sparitic flank facies and 3. loferitic capping facies. The central massive or crudely bedded part of the mounds consists of white or light grey clotted micrite. Macrofossils are rare. The sparitic flank facies in contrast consists of coarse and densely packed crinoidal wackestone-floatstones with some brachiopod shell debris. Solitary rugose corals, tabulate corals, stromato-poroids and fragments of mollusks are also abundant. The tops of the mounds are usually covered with loferitic pelmicrites or oolitic grainstone caps. Stromatactis-like structures are very rare and poorly developed in the South Ferganian mud mounds. However, almostin all such mounds horizons of calcitic breccias can be found. In order to explain all the features found in the Fergana mounds an ‘atoll-like’ model has been proposed which starts the evolution of the mud mounds with a small nucleus bioherm. The main stage of the evolution corresponds to an atoll-like structure developing on the surface of shallow water platforms. White clotted micrite of the mound core facies is interpreted as a accumulation of fine-grained sediment in an inner lagoon flanked by crinoidal bar deposits. The mound flank facies represents the atoll rim deposits from where the carbonate mud is derived. The capping loferitic facies is considered as tidal flat deposit that developed on top of the buildups during the last stage of its evolution. The knoll shape of the mounds is explained by the retreat of the atoll flanking crinoidal bars back into the inner lagoon during the rise in sea level. Stromatactis-like structures of small cavities filled with sparry calcite owe their existence to burrowing organisms. Calcitic breccias are interpreted as paleokarst collapse breccias. They indicate that the tops of the mud mound became subaerially exposed. Other evidence for a subaerial exposure can be seen in the occurrence of Variscian ‘black and white’ limestone gravel on the tops of some mud mounds. According toWard et al. (1970) these sediments were produced above the sea level at the edge of hypersaline lakes situated on islands.     

Organization of centromeres in the decondensed nuclei of mature human sperm

The localization of centromeres in mature human sperm was shown by immunofluorescent labeling and nonisotopic in situ hybridization. In the decondensed nucleus structural elements (dimers, tetramers, linear arrays and V shape structures) formed by individual centromeres of nonhomologous chromosomes were observed. They organize the compact chromocenter, which was shown for nuclei decondensed to a low extent. The chromocenter is buried inside the nucleus; in contrast, telomeric regions of chromosomes were tentatively localized on the periphery. Thus, a gross architecture, which can influence selective unpackaging of the paternal genome upon fertilization, exists in human sperm.     

The role of ecosystem transpiration in creating alternate moisture regimes by influencing atmospheric moisture convergence

The terrestrial water cycle links the soil and atmosphere moisture reservoirs through four fluxes: precipitation, evaporation, runoff, and atmospheric moisture convergence (net import of water vapor to balance runoff). Each of these processes is essential for sustaining human and ecosystem well-being. Predicting how the water cycle responds to changes in vegetation cover remains a challenge. Recently, changes in plant transpiration across the Amazon basin were shown to be associated disproportionately with changes in rainfall, suggesting that even small declines in transpiration (e.g., from deforestation) would lead to much larger declines in rainfall. Here, constraining these findings by the law of mass conservation, we show that in a sufficiently wet atmosphere, forest transpiration can control atmospheric moisture convergence such that increased transpiration enhances atmospheric moisture import and results in water yield. Conversely, in a sufficiently dry atmosphere increased transpiration reduces atmospheric moisture convergence and water yield. This previously unrecognized dichotomy can explain the otherwise mixed observations of how water yield responds to re-greening, as we illustrate with examples from China's Loess Plateau. Our analysis indicates that any additional precipitation recycling due to additional vegetation increases precipitation but decreases local water yield and steady-state runoff. Therefore, in the drier regions/periods and early stages of ecological restoration, the role of vegetation can be confined to precipitation recycling, while once a wetter stage is achieved, additional vegetation enhances atmospheric moisture convergence and water yield. Recent analyses indicate that the latter regime dominates the global response of the terrestrial water cycle to re-greening. Evaluating the transition between regimes, and recognizing the potential of vegetation for enhancing moisture convergence, are crucial for characterizing the consequences of deforestation as well as for motivating and guiding ecological restoration.     

Microwave influence on the isolated heart function: II. Combined effect of radiation and some drugs

The combined effects of microwave radiation and some drugs were studied in an isolated frog auricle preparation. The experiments established that exposure to pulse-modulated 915 MHz microwaves for up to 40 min had no effect on either the rate or the amplitude of spontaneous auricle twitches, unless the average absorbed power was high enough to produce preparation heating. Treatment of the preparation with saline containing (0.6–3.0) 10^?5 M of propranolol or (0.5–1.5) 10^?7 M of atropine altered neither its pacemaker nor its contractile functions; these drugs also had no effect when they were combined with nonthermal microwave irradiation. Caffeine (1 mM) strongly increased the average heart power, which was calculated as the product of twitch rate and amplitude. The caffeine effect appeared to be significantly augmented (by about 15%, P<0.02) under exposure to burst-type pulsed microwaves (pulse width, 1.5 msec; pause, 2.5 msec; 8 pulses/burst, 16 bursts/s; average SAR, 8–10 W/kg). By itself, this modulation was not effective; the heating of the preparation and saline during exposure was approximately 0.1°C, which could not account for the detected changes. The experimental results demonstrate that caffeine treatment increases the microwave sensitivity of the frog auricle preparation and reveals primarily subthreshold, nonthermal microwave effect. © 1995 Wiley-Liss, Inc.     

HSP70 Translocates into a cytoplasmic aggregate during lymphocyte activation

The percentage of T and B lymphocytes expressing a distinct cytoplasmic aggregate enriched in spectrin, ankyrin, and in several other proteins including protein kinase C greatly increases following various activation protocols. Members of the 70 kDa family of heat shock proteins (hsp70) temporarily bind to and stabilize unfolded segments of other proteins, a function apparently required for proper protein folding and assembly. Considering the multiprotein and dynamic nature of the lymphocyte aggregate, the possibility that hsp70 also might be associated with componets of this structure is considered here. Double immunofluorescence analysis indicates that hsp70 is a component of the lymphocyte aggregate and is coincident with spectrin in a subpopulation of freshly isolated, untreated lymphocytes from various murine tissues and in a T-lymphocyte hybridoma. When cell lysates of lymph node T cells are immunoprecipitated using an antibody against hsp70 or spectrin and then analyzed by Western blot utilizing the alternate antibody, it was found that hsp70 and spectrin coprecipitated with one another. Moreover, this coprecipitation could be abolished by addition of ATP. This latter observation was extended to lymphoid cells using a transient permeabilization procedure, and it was shown that addition of exogenous ATP results in the dissipation of the aggregate structure itself. Finally, conditions that result in T-cell activation and aggregate formation, i.e., treatment with the phorbol ester PMA or T-cell receptor cross-linking, also lead to the repositioning of hsp70 into the aggregate from a membrane/cytosolic locale in congruence with spectrin. These data suggest that hsp70 is an active component of the aggregate and that it may function in the interactions believed to occur in this unique activation-associated organelle. © 1995 Wiley-Liss, Inc.     

Structural features of archaebacterial and eukaryotic proteasomes

The 26S proteasome is the central protease of the ubiquitin-dependent pathway of protein degradation. The molecule has a molecular mass of approximately 2000 kD and has a highly conserved structure in eukaryotes. The 26S proteasome is formed by a barrel-shaped 20S core complex and two polar 19S complexes. The 20S complex has C2 symmetry and is formed by four seven-membered rings of which the outer rings (-type subunits) are rotated by 25.7° relative to the inner rings while the inner rings (-type subunits) are in register. From a comparison of the activity and regulation of the 26S and 20S particles it can be deduced that the 20S particle contains the protease activity while the 19S complex contains isopeptidase, ATPase and protein unfolding activities. In this article we describe the structures of various proteasome complexes as determined by electron microscopy and discuss structural implications of their subunit sequences.