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The analysis of the protein composition of exhaled breath to diagnose diseases of the respiratory system raises a problem of differentiation proteins of expressed in the tissues of the lungs and respiratory tract (endogenous) and got in the respiratory system from the ambient air in the process of respiration (exogenous). In this work an attempt was made to estimate a set of exhaled exogenic proteins by mass spectrometry coupled with nanoflow HPLC. Six-month isolation of healthy donors indoors with air cleaned of dust leads to removal from the spectrum of exhaled proteins of some keratins that are considered therefore to be exogenic. Non-keratin proteins may also circulate between the ambient air and human respiratory ways, but their concentration appears to be significantly lower the keratin concentrations (especially epidermis keratin). Among non-keratins dermcidin seems to be the most significant exogenic protein of exhaled air. The conclusion of the diagnostic value of exhaled proteins can be done only after careful comparison of the results of quantitative and qualitative analysis of their composition in norm and pathology for a statistically significant sample of donors.

ISSN 16076729, Doklady Biochemistry and Biophysics, 2011, Vol. 437, pp. 105–108. © Pleiades Publishing, Ltd., 2011.Original Russian Text © K.N. Kashkin, E.A. Musatkina, A.V. Komelkov, D.A. Sakharov, E.V. Trushkin, E.A. Tonevitsky, T.V. Vinogradova, E.P. Kopantzev, M.V. Zinovyeva,O.V. Kovaleva, K.A. Arkhipova, I.B. Zborovskaya, A.G. Tonevitsky, E.D. Sverdlov, 2011, published in Doklady Akademii Nauk, 2011, Vol. 437, No. 6, pp. 837–841.

Extracellular concentration of heat shock protein (Hsp) with a molecular weight of 70 kDa (Hsp70) rapidly increases in the serum in response to stress and returns to the basal level during recovery. Further regulation of its blood concentration is unclear. A possible regulator is HspBP1, a protein binding Hsp70. Binding to ATPase domain of Hsp70, HspBP1 inactivates it, thus acting as a factor of nucleotide exchange. Blood sera from athletes were examined at the beginning and end of the last mesocycle of the training period by two-staged immunoaffinity test system. The concentration of HspBP1 increased with decreasing Hsp70 concentration under conditions of long-term training. Presumably, the dynamics of Hsp70 and HspBP1 concentrations can serve as the test for evaluating the adaptation potential.

High and moderate intensity endurance exercise alters gene expression in human white blood cells (WBCs), but the understanding of how this effect occurs is limited. To increase our knowledge of the nature of this process, we investigated the effects of passing the anaerobic threshold (AnT) on the gene expression profile in WBCs of athletes. Nineteen highly trained skiers participated in a treadmill test with an incremental step protocol until exhaustion (ramp test to exhaustion, RTE). The average total time to exhaustion was 14:40 min and time after AnT was 4:50 min. Two weeks later, seven of these skiers participated in a moderate treadmill test (MT) at 80% peak O(2) uptake for 30 min, which was slightly below their AnTs. Blood samples were obtained before and immediately after both tests. RTE was associated with substantially greater leukocytosis and acidosis than MT. Gene expression in WBCs was measured using whole genome microarray expression analysis before and immediately after each test. A total of 310 upregulated genes were found after RTE, and 69 genes after MT of which 64 were identical to RTE. Both tests influenced a variety of known gene pathways related to inflammation, stress response, signal transduction and apoptosis. A large group of differentially expressed previously unknown small nucleolar RNA and small Cajal body RNA was found. In conclusion, a 15-min test to exhaustion was associated with substantially greater changes of gene expression than a 30-min test just below the AnT.

We report on the novel protocol for enzyme immobilization into gel of siloxanes using water-organic mixtures with the high content of organic solvent as a reaction medium. Hydrolysis of alkoxysilanes carried out without excessive dilution with water resulted in more active and stable enzyme containing membranes. Immobilization of an inherently labile lactate oxidase according to the proposed sol-gel protocol over Prussian Blue modified electrode resulted in an advanced lactate biosensor characterized with a sensitivity of 0.18 A M(-1) cm(-2) in the flow injection analysis (FIA) mode over a wide dynamic range. A comparison with the known sensors has shown that analytical performances of the elaborated lactate biosensor are advantageous over both published systems and commercialized devices. The biosensor shows an appropriate stability and is suitable for clinical analysis (including noninvasive diagnostics) and food quality control.

It was shown that GC rats predisposed to catalepsy do not differ from Wistar rats in success rate or latency (time of finding of the hidden escape platform) in the Morris water test. However, unlike Wistar, GC rats are inclined to passive drift and longer floating episodes. Rats of the MD+ strain predisposed to hyperkinesis in the form of horizontal pendulum-like movements of the head and upper limb girdle show longer latency and lower rate of successful trials than Wistar or MD-, rats selected for absence of the pendulum-like movements.

The efficiency of recognition of masked visual images (Arabic numerals) under conditions of listening to classical (intensity 62 dB) or rock music (25 dB) increased. Coherence of potential in the frontal cortical region characteristic of the masked image recognition increased under conditions of listening to music. The changes in intercenter EEG relations were correlated with the formation of "the recognition dominant" at the behavioral level. Such behavioral and EEG changes were not observed during listening to louder music (85 dB) and listening to music of other styles, however, the coherence between potentials of the temporal and motor areas of the right hemisphere increased, and the latency of hand motor reactions decreased. The results suggest that the "recognition dominant" is formed under conditions of establishment of certain relations between the levels of excitation in the corresponding centers. These findings should be taken into consideration in case if it were necessary to increase the efficiency of the recognition.

Intensive exercise triggers the cascade processes of body adaptation, including modulation of splisosome functioning, and can lead to modification of its activity and choice of alternative exons. We studied the effect of exercise of the maximum aerobic power on activation of transcription of genes involved in the splicing process. Short-term exercise resulted in a significant increase of mRNA expression of genes encoding proteins involved in the formation of precatalytic splisosome: DDX17, DDX46, HNRNPR, PRPF4B, and SRPK2. The role of the detected regulators in initiation of splisosome assembly under conditions of maximally intensive exercise is discussed.

Extracellular heat shock protein with molecular weight of 70 kDa is a signal molecule of the immune system. It is secreted by the peripheral blood, liver and muscle cells in response to physiological, thermal, and mental stresses. The main goal of our study was to compare the levels of expression of heat shock protein (70 kDa) matrix ribonucleic acid in leukocytes and serum concentrations of the protein before and after physiological stress. In order to solve this problem, we developed enzyme immunoassay of serum heat shock (70 kDa) protein concentration and a method for evaluating the expression of matrix ribonucleic acid of this protein in leukocytes by the real time PCR. The concentration of 70 kDa heat shock protein in the serum increased 1.7 times as a result of even a short-term highly intense physiological stress, while the expression of its matrix ribonucleic acid in leukocytes increased 1.5 times. The individual features determine the response to physiological stress. Probable sources of 70 kDa heat shock protein are discussed.

A 4-week study of adaptation reserves of the body was carried out during medium intensive exercise (medium intensive training: 60-80% threshold anaerobic metabolism). Two groups of athletes were singled out by the results of pulsometry analysis: with less than 20% work duration at the level above the 80% threshold anaerobic metabolism and with more than 20% work duration at the level above 80% threshold anaerobic metabolism. No appreciable differences between the concentrations of total testosterone, growth hormone, and cortisol before and after exercise in the groups with different percentage of anaerobic work duration were detected. In group 1 the concentrations of free testosterone did not change throughout the period of observation in comparison with the levels before training. In group 2, the level of free testosterone increased in comparison with the basal level: from 0.61+/-0.12 nmol/liter at the end of week 1 to 0.98+/-0.11 nmol/liter at the end of week 4 (p

To elucidate mechanisms that underlie the profound physiological effects of the monoamine precursors 5-hydroxy-L-tryptophan (5-HTP) and L-3,4-dihydroxyphenylalanine (L-DOPA), we examined their action on single monoaminergic neurons isolated from the ganglia of the gastropod snail Lymnaea stagnalis. In isolated serotonergic PeA motoneurons, 5-HTP produced excitation. The effect was mimicked by serotonin at 0.5-1 microM, masked by pretreatment with serotonin at higher concentrations, and abolished by the inhibitor of aromatic amino acid decarboxylase (AAAD) m-hydroxybenzylhydrazine (NSD-1015), the inhibitor of the vesicular monoamine transporter reserpine or the serotonin receptor antagonist mianserin. Exposure of the dopaminergic interneurons RPeD1 to L-DOPA caused a biphasic effect composed of a depolarization followed by a hyperpolarization. AAAD inactivation with NSD-1015, as well as the blockade of dopamine receptors with sulpiride, resulted in the enhancement of the excitatory effect, and the abolition of the inhibitory effect. Dopamine caused hyperpolarization and masked the inhibitory phase of L-DOPA action. The results show that precursors affect the rate of firing of isolated monoaminergic neurons and that their effect is completely or partially mediated by the enhanced synthesis of the respective neurotransmitter, followed by extrasynaptic release of the latter and activation of extrasynaptic autoreceptors.

A pharmacological model of hunting behavior, previously developed by the authors on the basis of the utilization of 5-hydroxytryptophan (5-HTP), a precursor of serotonin synthesis (5-HT), was employed in order to determine its cytochemical and neurophysiological correlates. In the presence of baseline behavior, 5-HT-containing neurons were not detected by the glyoxylate method, but they were readily identified in the buccal, cerebral, and pedal (the greatest number) ganglia of the CNS and in the peripheral organs during hunting behavior induced by an injection of 5-HTP (100-200 micrograms/g). The disappearance of pauses in the initial unsustained fictive swimming, an increase in the frequency of the rhythm of swimming and in the generation of spikes in each cycle, as well as a change in the phasing of the locomotor cycle, the neurophysiological correlates of "fast" locomotion in the presence of 5-HTP-induced hunting behavior, were recorded in isolated pedal ganglia in the presence of 5-HTP (100-200 micrograms/ml). The activity of 5-HT-neurons of the pedal ganglia, which was initially synchronized with the episodes of swimming, increases against the background of 5-HTP and becomes continuous. The effects of exogenous and endogenous 5-HT on the generator of locomotion are identical, and are induced by the direct action of the mediator on the membrane of the locomotor neurons. The hypothesis of the participation of 5-HT in the monoaminergic control of behavior is discussed.

The efficiency of the recognition of masked visual images (Arabic numerals) increased when accompanied by classical (62 dB) and rock music (25 dB). These changes were accompanied by increases in the coherence of potentials in the frontal areas seen on recognition without music. Changes in intercenter EEG relationships correlated with the formation a dominant at the behavioral level. When loud music (85 dB) and music of other styles was used, these changes in behavior and the EEG were not seen; however, the coherence of potentials in the temporal and motor cortex of the right hemisphere increased and the latent periods of motor reactions of the hands decreased. These results provide evidence that the “recognition” dominant is formed when there are particular ratios of the levels of excitation in the corresponding centers, which should be considered when there is a need to increase the efficiency of recognition activity in humans.

To detect neuroactive factors in biological fluids, we have previously elaborated a moveable biosensor, the preparation of isolated neuron penetrated with recording microelectrode [1]. Here, we report that, in the pond snail, signals emanated from a central pattern generator (CPG) for feeding and detected by such biosensor correspond to long-term changes in motor output of the CPG as recorded from buccal nerves. The results suggest that combined recording from motor nerves, neurons in situ and the neuron-based biosensors may be a useful tool in studies of the mechanisms underlying network organisation.

A novel excitatory higher-order input (named Input-HO) to the feeding network of the pond snail, Lymnaea stagnalis , as well as a reliable procedure that makes it possible to initiate and maintain the Input-HO, are described. The Input-HO is primarily targeted to two bilaterally symmetrical pairs of extrabuccal projecting interneurons, the serotonergic Cerebral Giant Cells (CGCs or C1) and FMRFamidergic Pleural-Buccal cells (PlBs). Judging from the results of transection experiments, Input-HO originates in the paired cerebral ganglia where its left and right sources coexist. These sources discharge independently or, sometimes, one after the other. Both inputs project to the ipsilateral CGC/PlB only. In turn, the activity of PlB decreases the frequency of Input-HO bouts. We suggest that the Input-HO represents a prominent part of the mechanism controlling feeding behavior in Lymnaea .

1. 1. Bathing the isolated CNS of A. fasciata in a 5-hydroxytryptophan (5HTP)-containing saline (50–120 μg/ml) induces sustained patterned rhythmic bursts in initially non-bursting neurones of the pedal ganglia. 2. 2. The neurones have been found within a specific area reportedly occupied by motor cells which supply locomotor muscles. 3. 3. The latency of the effect is 20–45 min and corresponds to that of actual swimming in whole animals injected with 5HTP. 4. 4. The frequency of bursts corresponds to that of cyclic swim movements. 5. 5. The rhythm is intrinsic to an, as yet unidentified, source of excitatory input. 6. 6. It is concluded from this evidence that metabolization of 5HTP results in release of a central program for locomotion, the fictitious swimming. 7. 7. Other 5HTP-induced changes include a considerable enhancement in firing of serotonin-containing neurones of the pedal ganglia. 8. 8. The 5HTP-treated isolated CNS is suggested to provide a favourable preparation to reveal the neuronal mechanisms of locomotor program generation in Aplysia .

The content of biogenic amines: dopamine, noradrenaline and serotonine, in rats of cataleptic strain GC as compared with the control strain Wistar at the age of 1 and 5 months is decreased, the maximal decrease being found in the so-called "nervous" animals. The aldosterone content was decreased at 5 month age in the GC rats. The testosterone content at the age of 1 month in GC rats does not differ from that in Wistar rats, but at the age of 5 months it was decreased as compared to Wistar, the maximal decrease being found again in "nervous" GC rats. The data obtained point to peculiarities of ontogenetic regulation and to commonness of mechanisms of catalepsy and "nervousness" in GC rats.

Changes in EEG coherence were studied in subjects without special musical education when they were listening to low classic music (below 30 dB). The numbers of significant changes in coherence function in the left and right hemispheres were different. Predominant increase in coherence in the alpha 2, beta 1, and beta 2 frequency ranges was observed in the right temporal-central and parietal-occipital cortical areas. The left-hemispheric coherence was predominantly, decreased on the account of its decrease in the alpha range.

The pleural interneuron PlB is a white neuron in the pleural ganglion of the snail Lymnaea. We test the hypothesis that it inhibits neurons at all levels of the feeding system, using a combination of anatomy, physiology and pharmacology. There is just one PlB in each pleural ganglion. Its axon traverses the pedal and cerebral ganglia, running into the buccal ganglia. It has neuropilar branches in the regions of the cerebral and buccal ganglia where neurons that are active during feeding also branch. Activation of the PlB blocks fictive feeding, whether the feeding rhythm occurs spontaneously or is driven by a modulatory interneuron. The PlB inhibits all the neurons in the feeding network, including protraction and retraction motoneurons, central pattern generator interneurons, buccal modulatory interneurons (SO, OC), and cerebral modulatory interneurons (CV1, CGC). Only the CV1 interneuron shows discrete 1:1 IPSPs; all other effects are slow, smooth hyperpolarizations. All connections persist in Ca(2+)/Mg(2+)-rich saline, which reduces polysynaptic effects. The inhibitory effects are mimicked by 0.5 to 100 micromol l(-1) FMRFamide, which the PlB soma contains. We conclude that the PlB inhibits neurons in the feeding system at all levels, probably acting though the peptide transmitter FMRFamide.

Single neurons (soma and proximal process) were isolated from the serotonergic (5-HT-ergic) PedA cluster in experiments on the pond snail Lymnaea stagnalis, and changes in the electrical activity of isolated neurons were observed during repeated movement of these cells towards and away from the surface of the CNS. The position of cell bodies of 5-HT-ergic neurons had excitatory effects on the isolated neuron. This effect was maximal (at 10(-8)-10(-7) M 5-HT) when neurons were brought close to the PedA cluster and were further enhanced by addition of the 5-HT precursor 5-hydroxytryptophan at concentrations of (1-2) x 10(-4) M. The results obtained here provide evidence 5-HT-ergic neurons cooperate during 5-HT-dependent behaviour, this being based on excitatory interactions at the level of cell bodies.

Analysis of ECGs recorded in 3 standard leads, 3 augmented limb leads, and 3 chest leads revealed typical signs of left ventricular hypertrophy with a relative deterioration of coronary blood supply in NISAG rats, a new strain with hereditary arterial hypertension. These signs are considered to be characteristic of an established arterial hypertension and may be taken as evidence that the NISAG strain can serve as an adequate animal model of human hypertensive disease.

In the cricket Gryllus bimaculatus, the opiate antagonist naloxone, 30 microg in 50 microl into hemolymph per animal, caused a release of intraspecific aggression in male-losers and in females. Naloxone had no significant effect on the aggression of winners and isolated males. The agonist of mu-opiate receptors DAGO, 45 microg, suppressed aggression in male winners and decreased the percentage and duration of contact fights between previously isolated males. The results suggest that, during social contacts, the activity of the endogenous opioid system may contribute to suppressing aggression in subordinate males, as well as in females.

Approach to regions occupied by perikarya of the snail 5-HT-ergic neurons produced excitation. The strongest action was recorded on the PedA cluster surface, and it was further enhanced by adding the 5-HT precursor. The findings suggest that the mechanisms underlying 5-HT-dependent behaviour include a mutual excitatory co-operation between somata of neighbouring 5-HT-ergic neurons.

We present the first steps in the elaboration of an approach of extracellular matrix-targeted local drug delivery (ECM-LDD), designed to provide a high concentration, ubiquitous distribution, and long-term retention of a drug within the vessel wall after local intravascular delivery. The approach is based on the concept of a bifunctional drug comprising a "therapeutic effector" and an "affinity vehicle," which should bind to an abundant component of the vessel wall. The aim of the present study was to select molecules suitable for the role of affinity vehicles for ECM-LDD and to study their intravascular delivery and retention ex vivo and in an animal model. By use of fluorescence microscopy, the following molecules were selected on the basis of strong binding to cross sections of human vessels: protamine, polylysine, polyarginine, a glycosaminoglycan-binding peptide from vitronectin, and a synthetic dendrimer. With polylysine as a prototypic affinity vehicle, we showed that after intravascular delivery, polylysine was concentrated throughout a luminal layer of the vascular wall to an extremely high concentration of 20 g/L and was retained therein for at least 72 hours of perfusion without noticeable losses. Low molecular weight (fluorescein) and high molecular weight (hirudin) compounds could be chemically conjugated to polylysine and were retained in the vessel wall after intravascular delivery of the conjugates. In conclusion, by use of the ECM-LDD method, an extremely high concentration and long-term retention of locally delivered drug can be reached. Polycationic molecules can be considered as potential affinity vehicles for ECM-LDD. Chemicals/CAS: Glycosaminoglycans; Hirudins; Peptides; polyarginine, 25212-18-4; Polylysine, 25104-18-1; Protamines; Vehicles

High-frequency ultrasound has been shown to accelerate enzymatic fibrinolysis. One of the supposed mechanisms of this effect is the enhancement of mass transport by acoustic streaming, i.e., ultrasound-induced macroscopic flow around the clot. In this study, which is aimed at further elucidating the mechanisms of the acceleration of fibrinolysis by ultrasound, we investigated whether ultrasound would accelerate fibrinolysis if the flow around the thrombus is already present, as may occur in vivo. The effect of the ultrasound-induced temperature rise was also studied. In a model of a plasma clot submerged in plasma, containing tissue-type plasminogen activator, mild stirring of the outer plasma producing a shear rate of 40 seconds(-1) at the surface of the clot resulted in a two-fold acceleration of lysis. A similar effect was obtained with ultrasound (1 MHz, 2 W/cm(2)). Furthermore, if ultrasound was applied together with stirring, only 30% acceleration by ultrasound was documented, fully attributable to the concomitant temperature rise. In a model with tissue-type plasminogen activator incorporated throughout a plasma clot, the effect of ultrasound (two-fold shortening of lysis time) was fully attributable to the concomitant temperature rise of a few degrees. We concluded that the acceleration of enzymatic plasma clot lysis by high-frequency ultrasound in the models we used can be largely explained by a combination of the effects of heating and acoustic streaming, equivalent to mild stirring. The thermal effects can hardly be utilized in vivo due to the danger of tissue overheat. The therapeutic advantage of transcutaneous high-frequency ultrasound as an adjunct to thrombolytic therapy may appear limited to the situations where there is no flow in the direct environment of the thrombus.

Administration of specific oligonucleotide selectively inhibiting alpha(2A)-adrenoceptor gene expression into the locus coeruleus of male rats for 3 days activated the hypothalamic-pituitary-adrenal system, which was manifested in a rise of blood plasma corticosterone content in rats with normal and hypertrophied (after castration) adrenal glands. These data indicate that alpha(2A)-adrenoceptors of the locus coeruleus are involved in the regulation of basal plasma corticosterone content.

Alkaloids (ergotamine and ergometrine) were shown to slow down rhythmic locomotion in even distantly related molluscs. The same is true for crawling pulmonate, pond snail Lymnaea stagnalis. The ergotamine activation of a powerful inhibitory input to pedal neurones involved in locomotor rhythmicity, was shown. The data obtained suggest that the conservative target for ergots is located outside rather than within the central pattern generator for locomotion.

1. In the cricket Gryllus bimaculatus, effects of opiate receptor antagonist naloxone, 9 or 30 microg per animal, on aggressive behavior were investigated. 2. Naloxone had no significant impact on aggression of isolated and dominant males. In contrast, the drug caused a dramatic release of social aggression in female and subordinate male crickets. 3. The results suggest that activity of the opioid system contributes to suppress aggression in subordinate males, as well as in females, during social contacts.

1. We have studied morphology, physiology and chemistry of a bilateral pair of pleural-to-buccal projecting neurons (PlB cells) of the pond snail Lymnaea stagnalis. Intracellular dye fills revealed axon arborization within neuropiles of ipsilateral pedal and cerebral ganglia, as well as in both buccal ganglia. Terminal axons of the left and right PlBs showed close proximity within the buccal commissure. 2. The left and right PlB neurons have been found electrotonically coupled and, sometimes, generating synchronous spikes. 3. The results show that two PlB cells operate as a single unit, and that paired buccal networks responsible for feeding rhythm are treated by the PlBs as a single target.

1. We have found that, in preparations of isolated CNS of the pond snail Lymnaea stagnalis, both serotonin (5HT) and dopamine (DA), as well as their respective precursors, 5HTP and DOPA, are effective in producing fictive intense (muscular) locomotion. 2. Phase-coupled to each of the above pedal rhythms are numerous identifiable pedal neurons including the respiratory interneuron RPeD1, thus suggesting interaction between networks responsible for locomotion and air breathing. 3. The novel DA/DOPA-dependent motor rhythm resembles the 5HT/5HTP-dependent one in terms of activity of identifiable pedal neurons, being however considerably slower than the latter. 4. The results of transection experiments suggest that each of the rhythms is generated by a paired CPG lying entirely within the pedal ganglia.

The effects of music with specific intensity on the latencies of the left or right hand motor responses to visual stimuli have been studied. When the latency of the initial motor response is more than 400 ms, the music accompaniment decreases the latency of the motor response of the left hand. It is supposed that the decrease in the mean latency of the left hand response in subjects who are not professional musicians is related to the activation effect of music on the right hemisphere. Music has no effect when the initial motor responses have shorter latencies.

We examined the effects of selective serotonin depletion and opioid ligands on social rank and related escape behavior of the cricket Gryllus bimaculatus. Establishment of social rank in a pair of males affected their escape reactions. Losers showed a lower and dominants a higher percentage of jumps in response to tactile cercal stimulation than before a fight. The serotonin-depleting drug alpha-methyltryptophan (AMTP) caused an activation of the escape reactivity in socially naive crickets. AMTP-treated animals also showed a lower ability to become dominants. With an initial 51.6+/- 3.6% of wins in the AMTP group, the percentage decreased to 26+/-1.6% on day 5 after injection. The opiate receptor antagonist naloxone affected fight and escape similarly as AMTP. In contrast to naloxone, the opioid agonist [d-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin decreased escape responsiveness to cercal stimulation in naive and subordinate crickets. We suggest that serotonergic and opioid systems are involved in the dominance induced depression of escape behavior.

The ionizing irradiation of rat fetuses during the last third of intrauterine development increased blood corticosterone level adulthood and decreased the open field locomotion of their adult offsprings of the next first nonirradiated generation. Treatment of the pregnant rats with glucocorticoids also decreased the offspring locomotion. Irradiation of fetuses in the middle of embryogenesis decreased blood corticosterone level in adulthood and shortened the open-field freezing reaction of their adult offsprings of the next first nonirradiated generation. Adrenalectomy of females before mating decreasing the blood corticosterone level had a similar effect on freezing duration of their adult offsprings. Irradiation of the ancestors within the last third of their intrauterine development had no effect on blood corticosterone level of their adult offsprings of the first generation and produced no behavioral alterations in their descendants of the next second nonirradiated generation. Irradiation of the ancestors in the middle of their embryogenesis decreased the stress-induced corticosterone response in their adult offsprings of the first generation and increased rearings and locomotion in their descendants of the next second nonirradiated generation. The data suggest that a single noxious treatment may have behavioral effects throughout two consequent generations of rats. Mother's glucocorticoid hormones may be one of the factors which transmit the effect.

In a number of cases, thrombolytic therapy fails to re-open occluded blood vessels, possibly due to the occurrence of thrombi resistant to lysis. We investigated in vitro how the lysis of hardly lysable model thrombi depends on the choice of the plasminogen activator (PA) and is accelerated by ultrasonic irradiation. Lysis of compacted crosslinked human plasma clots was measured after addition of nine different PAs to the surrounding plasma and the effect of 3 MHz ultrasound on the speed of lysis was assessed. Fibrin-specific PAs showed bell-shaped dose-response curves of varying width and height. PAs with improved fibrin-specificity (staphylokinase, the TNK variant of tissue-type PA [tPA], and the PA from the saliva of the Desmodus rotundus bat) induced rapid lysis in concentration ranges (80-, 260-, and 3,500-fold ranges, respectively) much wider than that for tPA (a 35-fold range). However, in terms of speed of lysis, these three PAs exceeded tPA only slightly. Reteplase and single-chain urokinase were comparable to tPA, whereas two-chain urokinase, anistreplase, and streptokinase were inferior to tPA. In the case of fibrin-specific PAs, ultrasonic treatment accelerated lysis about 1.5-fold. For streptokinase no acceleration was observed. The effect of ultrasound correlated with the presence of plasminogen in the outer plasma, suggesting that it was mediated by facilitating the transport of plasminogen to the surface of the clot. In conclusion, PAs with improved fibrin-specificity induce rapid lysis of plasminogen-poor compacted plasma clots in much wider concentration ranges than tPA. This offers a possibility of using single-or double-bolus administration regimens for such PAs. However, it is not likely that administration of these PAs will directly cause a dramatic increase in the rate of re-opening of the occluded arteries since they are only moderately superior to tPA in terms of maximal speed of lysis. Application of high-frequency ultrasound as an adjunct to thrombolytic therapy may increase the treatment efficiency, particularly in conjunction with fibrin-specific PAs.

Pedal cell RPeD1 of the pond snail L. stagnalis becomes involved in a central rhythm identified as an activity of the central pattern generator (CPG) for locomotion. The RPeD1 rhythm developed as driven by a synaptic input in isolated CNS preparations treated with 0.05 mM serotonin (5HT) or 0.1 mM 5-hydroxytryptophan (5HTP). The 5HT-induced co-ordinated rhythmic activity was retained by each of two pedal ganglia after complete isolation thus suggesting that the respective CPG lies entirely within the pedal portion of the CNS and is paired. The findings suggest that the RPeD1 switching from one network to another represents a neurotransmitter-dependent phenomenon.

Corticosterone levels in the blood and adrenal weights in adult rat males were increased after low-dose gamma irradiation during the last third of their intrauterine development; an increase of the dose decreased them. Decrease in testosterone levels and testis weights were dose-dependent. External and internal irradiation of females on the 11-14 days of pregnancy inhibited adrenals and increased testosterone levels and seminal vesicles weights in offspring. The changes of hormonal balance in adult animals depend on dose and period of ionizing irradiation during intrauterine development.

Current in vitro and animal tests for drug development are failing to emulate the systemic organ complexity of the human body and, therefore, to accurately predict drug toxicity. In this study, we present a multi-organ-chip capable of maintaining 3D tissues derived from cell lines, primary cells and biopsies of various human organs. We designed a multi-organ-chip with co-cultures of human artificial liver microtissues and skin biopsies, each a (1)/100,000 of the biomass of their original human organ counterparts, and have successfully proven its long-term performance. The system supports two different culture modes: i) tissue exposed to the fluid flow, or ii) tissue shielded from the underlying fluid flow by standard Transwell® cultures. Crosstalk between the two tissues was observed in 14-day co-cultures exposed to fluid flow. Applying the same culture mode, liver microtissues showed sensitivity at different molecular levels to the toxic substance troglitazone during a 6-day exposure. Finally, an astonishingly stable long-term performance of the Transwell®-based co-cultures could be observed over a 28-day period. This mode facilitates exposure of skin at the air-liquid interface. Thus, we provide here a potential new tool for systemic substance testing.