Using NADPH-diaphorase histochemistry, distribution of reactive neurons in the forebrain structures and motor cortex of rats was studied. Some reactive (NO-generating) neurons are associated with microvessels and, thus, can be involved in the regulation of regional blood flow. Almost ten years ago, it was found that an endothelium-derived relaxing factor (EDRF) is equivalent to nitric oxide (NO). For this finding, three American scientists, Robert F. Furchgott, Louis J. Ignarro, and Ferid Murad, were awarded the Noble Prize for Medicine in 1998. In the basal forebrain, there are four groups of cholinergic NO-generating neurons (Ch1-Ch4) projecting to the cortex. Thus, ascending pathways from the forebrain can also be involved in cortical activation through the release of NO [1,2]. Such corticopetal NO synthase (NOS)-containing cells and cholinergic networks in the cortex are implicated in several important brain phenomena, including arousal, attention, cortical plasticity, and memory. Abnormalities of the L-arginine-NO synthesis pathways in various structures of the basal forebrain and cortex are associated with severe impairment of cerebral functions. Dementia related to Alzheimer's disease results to a significant extent from a loss of the NADPH-diaphorase/NOS activity in the cortex [3][4][5]. It should be emphasized that colocalization of NOS immunoreactivity and NADPH reactivity has been demonstrated in a number of brain neurons. Cholinergic innervation of the cortex has been extensively studied. In humans, its main center of origin was identified as the nucleus basalis of Meynert; its equivalent in the rat is the substantia innominata (SI). Electron microscopic studies demonstrated that basal forebrain neurons project preferentially to the cortical pyramidal and NOScontaining non-pyramidal neurons; however, there are also projections to intraparenchymal arterioles, microvessels, and even capillaries. A significant proportion of these projections are cholinergic or GABA-ergic; in addition, they are NADPH diaphorasereactive [6,7].Based on the data on the co-localization of GABA and NADPH diaphorase (NADPH-d) in a population of cortical non-pyramidal neurons, or on that of acetylcholine and NADPH-d in a subpopulation of basal forebrain neurons, we suggested that these cells and their fibers and dendrites can be implicated in structural associations with the arterioles and microvessels.All experiments were carried out in accordance with the European Communities Council Directive of November 24, 1986 (86/609/EEC). Male Wistar rats (n = 8) weighing 250-300 g were used in the study. Frontal frozen sections of the brain (40 µm thick) were cut. The NADPH-d reactivity was detected according to a slightly modified standard histochemical method; for intensification of the reaction, disodium salt of malic acid (1.2 mg/ml; Sigma, USA) was added to the staining solution [8].The NADPH-d reactivity and distribution density of reactive neurons in the limbic structures and primary motor cortex were estimated. The neurons were ...

c-fos gene expression in the cervical spinal cord and amygdala was examined in anaesthetized rats following muscle fatigue caused by intermittent high-rate (100 s(-1)) electrical stimulation of the dorsal neck muscles (m. trapezius and m. splenius). Fatigue-related increases in c-fos expression were observed on the stimulated muscle side in the cervical C2-C4 (layers 1, 3-5, 7 and 10) spinal segments, bilaterally in the lumbar L4-L6 (layer 1) segments and in contralateral central (Ce), medial (Me), and basomedial (BM) amygdaloid nuclei. A scarce number of staining cells were found within lateral and basolateral nuclei. The rostro-caudal extent of c-fos expression in the spinal cord supports functional coupling of the cervical and lumbar regions during the neck muscle fatigue development. The distinct c-fos expression in the Ce and Me amygdaloid nuclei suggests that they may contribute to mediating the neck muscle fatigue-related nociception, autonomic and behavioural responses.

A comparative study of mmunoreactivity with respect to c-Fos protein in the motor (zones М1 and М2), medial prefrontal (PrL and IL), and cingular (Cg1 and Cg2) cortices allowed us to find significant differences between the intensities of expression of gene c-fos in these cortical regions in control rats (group 1) and animals trained to perform catching of food globules by the forelimb (i.e., realizing an operant food-procuring reflex, group 2). The density of distribution of Fos-immunoreactive (Fos-ir) neurons in rats of group 2 in motor and limbic cortical zones at +2.2 to +0.2 levels rostrally from the bregma were significantly lower than in control rats (Р < 0.05). In animals of group 2, we also found significantly greater numbers of Fos-ir neurons in the contralateral (with respect to the active extremity) zones of the cortex at all examined levels. These changes are probably related to functional changes in the cortex resulting from learning of motor habits in the course of training sessions for stabilization of the operant reflex. Histochemical estimation of the NADPH-diaphorase (NADPH-d) activity in the motor and limbic cortex showed that, in rats of both groups, the maximum number of labeled interneurons per slice in the М1 zone were observed in layers II/III, V, and VI (5.6 ± 0.4, 6.4 ± 0.5, and 14.0 ± 0.8, respectively, within 200 × 200 μm 2 areas). In the limbic cortex, NADPH-d-reactive (NADPH-d-r) interneurons were also met in layers II/III, V, and VI. Cortical NADPHd-r neurons with the Fos-ir nuclei were not found. The presence of spatial associations of the somata or processes of NADPH-d-r neurons with intraparenchimal arterioles and microvessels was a typical feature of the distribution of NADPH-d-reactivity in the М1 and М2 zones, as well as in Cg1, Cg2, PrL, and IL. The following succession of the density of neurovascular associations was observed: Cg1 > Cg2 > М1 > М2 > > PrL. As is supposed, NADPH-d-r neurons (i.e., cells generating NO) are involved in the control of regional blood flow in the studied cortical regions.

Background: Today, cardiovascular diseases cause 47% of all deaths among the European population, which is 4 million cases every year. In Ukraine, CAD accounts for 65% of the mortality rate from circulatory system diseases of the able-bodied population and is the main cause of disability. The aim of this study is to develop a medical expert system based on fuzzy sets for assessing the degree of coronary artery lesions in patients with coronary artery disease. Methods: The method of using fuzzy sets for the implementation of an information expert system for solving the problems of medical diagnostics, in particular, when assessing the degree of anatomical lesion of the coronary arteries in patients with various forms of coronary artery disease, has been developed. Results: The paper analyses the main areas of application of mathematical methods in medical diagnostics, and formulates the principles of diagnostics, based on fuzzy logic. The developed models and algorithms of medical diagnostics are based on the ideas and principles of artificial intelligence and knowledge engineering, the theory of experiment planning, the theory of fuzzy sets and linguistic variables. The expert system is tested on real data. Through research and comparison of the results of experts and the created medical expert system, the reliability of supporting the correct decision making of the medical expert system based on fuzzy sets for assessing the degree of anatomical lesion of the coronary arteries in patients with various forms of coronary artery disease with the assessment of experts was 95%, which shows the high efficiency of decision making. Conclusions: The practical value of the work lies in the possibility of using the automated expert system for the solution of the problems of medical diagnosis based on fuzzy logic for assessing the degree of anatomical lesion of the coronary arteries in patients with various forms of coronary artery disease. The proposed concept must be further validated for inter-rater consistency and reliability. Thus, it is promising to create expert medical systems based on fuzzy sets for assessing the degree of disease pathology.

Using the corresponding techniques, we visualized Fos-immunoreactive (Fos-ir) and NADPH diaphorasereactive (NADPH-dr) neurons in the medullary centers of the autonomic nervous system (ANS) of rats, which performed repetitive operant movements (catching of food globules from the manger by the left forelimb under conditions of high food motivation). Animals were trained to perform operant movements in 30 min-long everyday sessions during 12 days. The duration of a single food-procuring movement was about 600 msec. Realization of the operant reflex was accompanied by clearly expressed motivational/affective reactions. The heart rate (HR) in the course of each operant movement sharply dropped (on the 10th day of training, by 12%, on average) with subsequent recovery of this parameter within 3-4 sec. In the course of 30-min-long training sessions, the mean HR gradually decreased (in the examined group, 7%, on average) within an interval from the 5th to the 20th min with subsequent recovery until the end of the training session. The mean numbers of Fos-ir neurons in the medullary nuclei of the ANS (Sol, IRt, СVL, RVL, Amb, 10, and MdD) of rats performing food-procuring movements (n = 4) were significantly (Р < 0.05) greater than those in the control, and the intensities of c-fos expression in these structures corresponded to the following succession: Sol > IRt > СVL+RVL/СVL > RVL. Large Fos-ir neurons were observed in the dorsal motor nucleus of the n. vagus (10) and in the Amb/RAmb nuclei. In a considerable proportion of neurons of the Sol and single cells of the 10 and Amb, we observed double labeling (Fos-ir + NADPH-dr). Thus, operant food-procuring movements are accompanied by episodes of bradycardia related to each separate realization; in addition, a long-lasting tonic decrease in the HR developed. These autonomic reactions are mediated by the abovementioned medullary ANS nuclei. It is supposed that the respective weakening of inhibitory sympathetic effects on spindle receptors of the muscles involved in realization of the above operant movements can provide facilitation of generation of proprioceptive impulsation, facilitation of monosynaptic spinal reflexes, and, finally, an increase in the efficacy of targeted limb movements directed toward food procurement.