Effect of thermal treatment on the structure and biological properties of tuberculin

The effect of heat treatment used in the course of tuberculin production by Mycobacteria tuberculosis cells on the structure and biological activity of tuberculin was studied. Methods of preparative and analytical gel-filtration were used. The largest quantity of the biologically active component was obtained after 2-hour heat treatment of the mycobacteria culture.     

Micellar electrokinetic chromatography with polyelectrolyte complexes as micellar pseudo-stationary phases

The separation of dansyl (DNS-AAs) and carbobenzoxy (CBZ-AAs) amino acids using micellar electrokinetic chromatography employing polyelectrolyte-surfactant complexes (PSC) formed in the reaction between polyacrylic acid (PAA) and dodecyltrimethylammonium bromide (DTAB) as pseudo-stationary phases was described. The PSCs were stabilized by hydrophobic interactions of alkyl chains of the surfactant ions and converted to an intramolecular micellar-like phase. The running buffer was a 50mM solution of sodium phosphate (pH 6.0) containing 4.6-20.2mM PSC, in which a part of carboxyl groups of PAA was blocked by aliphatic amines. For the systems with 7.9mM of PAA/DTAB complex (phi=0.30, phi-composition of water-soluble polyelectrolyte complex) as a pseudo-stationary phase, the peaks of six dansyl amino acids (DNS-AAs) were baseline resolved. The separation in this case is based on a complex distribution mechanism of the dansyl derivatives between the free buffer and the intramolecular micellar-like phase of the water-soluble PSC. On the other hand, the additives of PAA/DTAB complex (phi=0.30) to the running buffer does not essentially affect on the electrophoretic behaviour of the CBZ-AAs, the variant MEKC is not realized. The influence of the concentration of the complex of PAA/DTAB on the electrophoretic behaviour of analytes was investigated. Relative retentions and relative selectivities were used for describing electrophoretic behaviour of the amino acid derivatives.     

Changes in the Balance between Membrane-Bound and Soluble Forms of CD95 (Fas) during Selection of Tumor Cells in vivo

Studies concerning the expression of the receptor CD95 (Fas) by tumor cells and the role of this protein in apoptosis induced by the effector host cells that bear Fas-ligand are mainly focused on the membrane-bound form of Fas. There are only a few data about the production of the soluble form of Fas by the tumor cells, its role in the interaction with the effector host cells, and the possible changes in the synthesis of this protein during tumor progression. In the present work, three in vivo transformed parental cell lines of different origin and 24 of their variants isolated after a short cycle of natural selection in vivo were studied. It was demonstrated for the first time that: 1) production of the soluble Fas by all selected in vivo variant tumor cell lines increased significantly (2-10-fold) in comparison to the initial (parental) cell lines and did not depend on the origin of the parental lines. At the same time, the expression of the membrane bound form of Fas decreased considerably; 2) variations of the balance between membrane-bound and soluble forms of Fas in selected in vivo variant cells and the expression of the [H₂O₂ ^CA + PGE^S]-phenotype by these cells (this phenotype determines one of the essential mechanisms of the protection of a tumor cell in vivo) possibly represent independent secondary changes acquired during tumor progression in vivo.     

Neuroprotection in epilepsy

Brain damage and neuronal loss caused by traumatic brain injury, ischemic stroke, and symptomatic status epilepticus can lead to severe long-term consequences, such as impairment in learning and memory and cognitive functions, and development of chronic epilepsy. This can be the result of morphologic and functional changes underlying temporal lobe epilepsy. Epilepsy patients have increased risk of status epilepticus. It is a life-threatening condition when seizures last for more than 30 min and trigger processes leading to neuronal apoptosis and necrosis in various parts of brain. Administration of neuroprotective drugs preventing these pathologic processes could improve the prognosis for such patients. However despite of active research of neuroprotective drugs, the effective ways to prevent brain damage resulting from prolonged seizures are yet to be found. Studies of neuroprotective properties of classic and novel anticonvulsant drugs showed that most of them do not have the sufficient neuroprotective effect and are not able to prevent epileptogenesis. Thus the studies of other potential neuroprotective drugs seem to be promising.     

Estimation of localization and dipole moment of alpha- and theta-rhythm sources by cluster analysis in healthy subjects and schizophrenics

In 12 healthy subjects and 9 schizophrenic patients in the background conditions (with eyes closed) EEG was recorded from 16 standard derivations (10-20 system) during 3 min. The record underwent the spectral analysis detecting alpha- and theta-frequency bands. After the preliminary narrow band filtration for the main frequencies the sources of the spontaneous rhythms were localized. The data on localization for all healthy subjects and patients were summarized. The K-means clustering was used for identification of the sources clusters which were revealed in occipital and parietal lobes and limbic cortex for alpha-rhythm and also in frontal, temporal and parietal regions, limbic cortex and hippocampus for theta-rhythm. In schizophrenic patients in comparison with healthy subjects there was revealed significant increase of the numbers of dipole sources of alpha-rhythm in the clusters localized in limbic cortex and hippocampus. For theta-rhythm there was significant increase of the dipole moment of the sources in the clusters localized in the temporal and frontal cortices and hippocampus in patients in comparison with the norm.     

Effect of anesthetics on efficiency of remote ischemic preconditioning

Remote ischemic preconditioning of hind limbs (RIPC) is an effective method for preventing brain injury resulting from ischemia. However, in numerous studies RIPC has been used on the background of administered anesthetics, which also could exhibit neuroprotective properties. Therefore, investigation of the signaling pathways triggered by RIPC and the effect of anesthetics is important. In this study, we explored the effect of anesthetics (chloral hydrate and Zoletil) on the ability of RIPC to protect the brain from injury caused by ischemia and reperfusion. We found that RIPC without anesthesia resulted in statistically significant decrease in neurological deficit 24 h after ischemia, but did not affect the volume of brain injury. Administration of chloral hydrate or Zoletil one day prior to brain ischemia produced a preconditioning effect by their own, decreasing the degree of neurological deficit and lowering the volume of infarct with the use of Zoletil. The protective effects observed after RIPC with chloral hydrate or Zoletil were similar to those observed when only the respective anesthetic was used. RIPC was accompanied by significant increase in the level of brain proteins associated with the induction of ischemic tolerance such as pGSK-3β, BDNF, and HSP70. However, Zoletil did not affect the level of these proteins 24 h after injection, and chloral hydrate caused increase of only pGSK-3β. We conclude that RIPC, chloral hydrate, and Zoletil produce a significant neuroprotective effect, but the simultaneous use of anesthetics with RIPC does not enhance the degree of neuroprotection.     

Neuronal correlates of the activation of the brain-reinforcing system in dogs

In analysis of neuronal activity of g. proreus of dogs brain in response to positive and differentiation conditioned cutaneous stimuli, in 44% of 78 studied neurons "late responses" appeared in the form of impulse activity intensification elicited by a positive conditioned stimulus after the end of eating and by a differentiation one--during conditioned signal action; it persisted in different neurones for 30--60 s. By means of various functional tests it has been shown that these reactions are a neurophysiological correlate of animals emotional state gradient.     

Unit activity of the prefrontal cortex during delayed alternation performance in monkey.

Unit activity in the dorsolateral regions of the prefrontal cortex (s. principalis) of the monkey was analysed by multineuronal recording technique. The sequence during delayed-response alternation included anticipatory stimulus, non-specific expectancy, conditioned cue, delay, trigger stimulus and alternation performance. Food reward completed the sequence. Unit activity in s. principalis was found to be involved in such learning processes as integration of behavioural acts into an accomplished programme and storage of traces in short-term memory. The latter can be observed in successive involvement of neuronal populations during a 10 sec delay as well as in rearrangements of unit activity, the maximum of which shifts to the latest part of the delay period. Unit activity in s. principalis reflects the level of correct response, corresponding correlates of which depend on the level of attention. The dorsolateral prefrontal cortex has structural and functional characteristics for short-term storage of external signals in its neuronal nets according to the level of attention.     

New-generation Skulachev ions exhibiting nephroprotective and neuroprotective properties

A mitochondria-targeted chimeric compound consisting of a rhodamine derivative linked to a plastoquinone molecule (10-(6′-plastoquinonyl)decylrhodamine, SkQR1) was studied under conditions of acute brain or kidney damage. A protective effect of this compound was demonstrated in a model of focal brain ischemia, rat kidney ischemia/reperfusion, myoglobinuria (rhabdomyolysis, or crush syndrome), and pyelonephritis. We found that a single intraperitoneal injection of SkQR1 diminishes the size of the ischemic zone in the brain and improves performance of a test characterizing neurological deficit in ischemic animals. An analog of SkQR1 not containing plastoquinone (C12R19) was not neuroprotective. The data show that SkQR1 is a nephroprotectant and neuroprotectant, which can be due to the antioxidative action of this Skulachev cation.