Changes in direct current input resistance in horizontal cells of fish retina during excitation

The input resistance of pike retinal horizontal cells was measured by means of coaxial electrodes under various conditions of illumination. With moderate intensities of illumination, the resistance (determined from a potential drop caused by the current passed through the microelectrode) increases, whereas at high saturating intensities it decreases, as compared with its value in darkness. Such changes in resistance of the horizontal cells explain the effects of input signals interaction in these cells, such as enhancement and complete saturation, observed earlier. Some properties of the horizontal cell response permit us to assume that the "active" cell response to polarization makes a substantial contribution to the measured resistance of these cells. Possible mechanisms of such changes in input resistance of horizontal cells are discussed.Institute of Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 3, No. 2, pp. 210–216, March–April, 1971.     

Fluorescent Labeling Preserving OCP Photoactivity Reveals Its Reorganization during the Photocycle

Orange carotenoid protein (OCP), responsible for the photoprotection of the cyanobacterial photosynthetic apparatus under excessive light conditions, undergoes significant rearrangements upon photoconversion and transits from the stable orange to the signaling red state. This is thought to involve a 12-Å translocation of the carotenoid cofactor and separation of the N- and C-terminal protein domains. Despite clear recent progress, the detailed mechanism of the OCP photoconversion and associated photoprotection remains elusive. Here, we labeled the OCP of Synechocystis with tetramethylrhodamine-maleimide (TMR) and obtained a photoactive OCP-TMR complex, the fluorescence of which was highly sensitive to the protein state, showing unprecedented contrast between the orange and red states and reflecting changes in protein conformation and the distances from TMR to the carotenoid throughout the photocycle. The OCP-TMR complex was sensitive to the light intensity, temperature, and viscosity of the solvent. Based on the observed Förster resonance energy transfer, we determined that upon photoconversion, the distance between TMR (donor) bound to a cysteine in the C-terminal domain and the carotenoid (acceptor) increased by 18 Å, with simultaneous translocation of the carotenoid into the N-terminal domain. Time-resolved fluorescence anisotropy revealed a significant decrease of the OCP rotation rate in the red state, indicating that the light-triggered conversion of the protein is accompanied by an increase of its hydrodynamic radius. Thus, our results support the idea of significant structural rearrangements of OCP, providing, to our knowledge, new insights into the structural rearrangements of OCP throughout the photocycle and a completely novel approach to the study of its photocycle and non-photochemical quenching. We suggest that this approach can be generally applied to other photoactive proteins.     

Increase of seizure threshold and resistance to stress in rats after immunization against fragment of diazepam-binding inhibitor]

White rats were immunized against fragment of diazepam-binding inhibitor octadecaneuropeptide (ODN) with conjugate ODN bovine serum albumin. This rats have reduced reactions of fear and anxiety in stress model of "open field" and in conflict Vogel test; their pain sensitivity ("tail flick" test) was lowered. The number and intensity of generalized seizure reactions after injection of pentylenetetrazole were decreased. The results show that active immunization to endogenous ODN has stress--protective and anti-seizure effects.     

Low-voltage underwater electric discharges: Physical properties and application possibilities

Electrical discharges that burn inside an electrolyte bulk and have challenging practical applications are presented. The data on electrophysical and spectral properties of an underwater end-face discharge allowed suggesting the scenario of the discharge origination and development. It includes three stages: bubble growth, a quiet discharge, and an active discharge stage.     

Detection of nanodiamonds in biological samples by EPR spectrometry

In model experiments in vitro, the applicability of the EPR spectrometry method for the detection of modified nanodiamonds (MNDs) in blood and homogenates of mouse organs has been established. A characteristic signal (g = 2.003, ΔH ≈ 10 G) is observed in the samples of biomaterials containing MNDs, the intensity of which increases linearly with the concentration of nanoparticles in the range of 1.6–200 μg MNDs per 1 mL of the sample. The EPR method in biomaterials reveals the presence of intrinsic paramagnetic centers, signals from which are superimposed on the signal from the MNDs. However, the intensity of these signals is small, which makes it possible to register the MNDs using EPR spectrometry with the necessary accuracy. The data obtained open up the prospects of using the EPR method for studies of the interorgan distribution, accumulation, and elimination of MNDs during their intravenous injection into experimental animals.     

A Method to Simultaneously Detect Changes in Intracellular Ca<Superscript>2+</Superscript> Concentration and Cell Volume

A method to simultaneously assess the changes in intracellular calcium concentration and cell volume in single cells was developed using the Ca²⁺-sensitive fluorescent probe Fura-2 and a three-dimensional image-surface reconstruction technique, respectively. Studies with this method showed that Fura-2 loading had no significant effect on the kinetics of A549 human epithelial cell swelling in a hypotonic solution, as well as the volume restoration kinetics. Significant changes in intracellular Ca²⁺ concentration were not observed in the examined volume modulation range. The results suggest that Ca²⁺-mediated signaling pathways are not involved in the autoregulation of the cell volume in A549 cells exposed to hypotonic conditions.     

Alterations of Antioxidant and Oxygen Transport Properties of Blood in Adenocarcinoma-Bearing Mice during Chemotherapy

The influence of murine intestinal adenocarcinoma on the antioxidant status and hemoglobin content in the blood was analyzed. Murine adenocarcinoma was accompanied with a decrease of the nonprotein thiol concentration and an increase in ceruloplasmin. A conformational change in the globin molecule but not in the heme group was revealed in the hemoglobin molecule. The changes in the protein appear to be caused by nonspecific damage that results from the processes of peroxidation.     

Linkage disequilibrium and haplotypes of the rs1042522, rs1625895, and rs1787862 markers of TP53 in patients with diffuse large B-cell lymphoma

The association of the rs1042522, rs17878362, and rs1625895 polymorphisms of TP53 with risk of non-Hodgkin’s lymphoma (NHL) has not been studied in full detail. NHL has many variants, and each individual polymorphism exerts only a minor effect, requiring the polymorphisms to be studied for particular histological subtypes of lymophomas and as components of haplotype groups. The objective of this work was to analyze the frequencies and linkage disequilibrium for rs1042522, rs1625895, and rs17878362 and their combined haplotype in patients with diffuse large B-cell lymphoma (DLBCL) and control subjects. Differences in linkage disequilibrium structure were observed for rs17878362, rs1042522, and rs1625895 in the Siberian population. The haplotype proved to be more informative than the individual TP53 polymorphisms in a case-control association analysis in DLBCL. Haplotype wArgG was associated with predisposition to DLBCL, while haplotypes wProG and dupProG were found to exert a protective effect. The effect of the haplotype at three TP53 polymorphisms was observed to vary depending on their homozygous or heterozygous state.     

Seasonal changes in the content of photosynthetic pigments in perennial grasses of cryolithic zone

Seasonal changes in the content of photosynthetic pigments and their ratio were studied in perennial gramineous (Psathyrostachys juncea Tzvel., Elymus sibiricus L., Bromopsis inermis Leys) and equisetaceous (Equisetum variegatum Schleich. ex Web., E. scirpoides Michx.) plants grown under conditions of cryolithic zone in Central and North-East Yakutia (the Pole of Cold). Simultaneously with growth in chlorophyll (Chl) content in all the studied species of perennial grasses of Central Yakutia, a considerable increase in the content of carotenoids (Car) of the xanthophyll cycle (XC) pigments was observed with the beginning of autumn temperature fall. Similar dependence was found in horsetails of the Pole of Cold region: the content of Car was increased by 1.2–3.0 times comparing to the summer values, whereas at the same time the concentration of oxygen-containing forms increased and the ratio of Chl to xanthophylls and Car decreased. In general, horsetails had typically a lower content of Chl and Car, which was determined not only by extremely harsh climatic conditions of the region but also by specific characteristics of Equisetaceae plants. Closer to winter time, the formation of secondary Car, rhodoxanthin, was also observed in the studied horsetail species. In this case, rhodoxanthin presumably plays more important antioxidant role than XC, which can not operate at constantly low temperatures. The obtained data suggest that one of the characteristics of cold hardening of perennial grasses under conditions of cryolithic zone of Yakutia is, presumably, the high level of primary, as well as secondary, Car with most profound antioxidant properties.     

Non-photochemical fluorescence quenching in photosystem II antenna complexes by the reaction center cation radical

In direct experiments, rate constants of photochemical (k_P) and non-photochemical (k_P⁺) fluorescence quenching were determined in membrane fragments of photosystem II (PSII), in oxygen-evolving PSII core particles, as well as in core particles deprived of the oxygen-evolving complex. For this purpose, a new approach to the pulse fluorometry method was implemented. In the “dark” reaction center (RC) state, antenna fluorescence decay kinetics were measured under lowintensity excitation (532 nm, pulse repetition rate 1 Hz), and the emission was registered by a streak camera. To create a “closed” [P680⁺Q_A^–] RC state, a high-intensity pre-excitation pulse (pump pulse, 532 nm) of the sample was used. The time advance of the pump pulse against the measuring pulse was 8 ns. In this experimental configuration, under the pump pulse, the [P680⁺Q_A^–] state was formed in RC, whereupon antenna fluorescence kinetics was measured using a weak testing picosecond pulsed excitation light applied to the sample 8 ns after the pump pulse. The data were fitted by a two-exponential approximation. Efficiency of antenna fluorescence quenching by the photoactive RC pigment in its oxidized (P680⁺) state was found to be ～1.5 times higher than that of the neutral (P680) RC state. To verify the data obtained with a streak camera, control measurements of PSII complex fluorescence decay kinetics by the single-photon counting technique were carried out. The results support the conclusions drawn from the measurements registered with the streak camera. In this case, the fitting of fluorescence kinetics was performed in three-exponential approximation, using the value of τ₁ obtained by analyzing data registered by the streak camera. An additional third component obtained by modeling the data of single photon counting describes the P680⁺Pheo^– charge recombination. Thus, for the first time the ratio of k_P⁺/k_P = 1.5 was determined in a direct experiment. The mechanisms of higher efficiency for non-photochemical antenna fluorescence quenching by RC cation radical in comparison to that of photochemical quenching are discussed.