[Role of nitric oxide in treatment of acute lung injury after surgery with extracorporeal circulation]. / Uloga azotnog oksida u lecenju akutnih plucnih ostecenja posle operacija izvedenih u uslovima vantelesnog krvotoka.

Postoperative acute lung injury (ALI) compromises oxygen transfer across alveolar-capillary membrane with consecutive hypoxia, one of its indicators being reduction of oxygenation index PaO2/FiO2 below 40 kPa (300 mm Hg). Management of ALI includes different procedures like mechanical lung ventilation (MLV), drugs and others. One of the new possibilities for treatment of ALI is nitric oxide (NO) inhalation. The aim of this prospective study was to examine the role of NO inhalation in treatment of ALI. 14 patients with ALI developed immediately after operation with extracorporeal circulation (ECC) were included in the study. Group A (n = 8) inhaled NO and group B (n = 6) did not inhale NO during treatment of ALI. All other therapeutic measures were the same in both groups. The groups were similar in relation to demographic data, type of surgery and duration of ECC. PaO2/FiO2 was calculated before operation (T1), immediately after surgery (T2) and after lung recovery, when the need for MLV stopped (T3). The duration of MLV was also registered. PaO2/FiO2 (kPa) in referent times was in group A 54.9 +/- 1.6, 33.8 +/- 1.2 and 46.2 +/- 0.8 and in group B 52.2 +/- 1.1, 33.5 +/- 1.5 and 47.3 +/- 0.9, respectively. There was a statistically significant decrease of PaO2/FiO2 in T2 and T3 vs T1 in both groups (p < 0.05), while the difference between the groups was not statistically significant. The duration of MLV (h) in group B (28.5 +/- 1.6) was statistically significantly shorter than in group A (63.1 +/- 8.7) (p < 0.01). According to the results of this study we conclude that NO inhalation during ALI after surgery with ECC significantly reduces the duration of MVL and improves pulmonary recovery.

Presumed 'prefrontal cortex' lesions in pigeons: effects on visual discrimination performance.

The posterodorsolateral neostriatum (PDLNS) in pigeons may be an equivalent of the prefrontal cortex (PFC) in mammals. Here we report that lesions of this brain region in pigeons have a detrimental effect on various learned visual discriminations. Pigeons with lesions of the overlying area corticoidea dorsolateralis (CDL) served as controls. Both the postoperative re-learning to criterion of a preoperatively learned simultaneous double visual mirror pattern discrimination and the learning of a simple successive go, no-go discrimination were impaired by the PDLNS lesions. The PDLNS and CDL groups did not differ significantly in the postoperative learning of a reversal of the simultaneous discrimination. The results are discussed in relation to the presumed equivalence between the avian PDNLS and the mammalian PFC.

Memory of radial maze behavior in pigeons after ablations of the presumed equivalent of mammalian prefrontal cortex.

Pigeons were trained and postoperatively tested in an 8-arm radial maze in which 1 arm was always used for start, 3 were never baited, and 4 were always baited. Of 2 groups of pigeons, 1 (n = 6) received ablations of the postero-dorso-lateral neostriatum (PDLNS) as well as the corticoid, and the other (n = 8) was sham operated. After the surgery, in the PDLNS group the number of reference memory errors (entering the never-baited arms) was significantly increased, but the number of working memory errors (entering previously visited baited arms) was not. Two of 6 pigeons with PDLNS ablations did not show any impairment, 3 were impaired in the reference memory, and 1 was impaired in working memory. This outcome resembles behavioral effects obtained in rats with prefrontal lesions. In pigeons and rats, the lesion seems to "release" the normal "win-shift" tendency and/or impair the ability to choose correctly in simultaneous multiple-choice situations.

Representation of a single vibrissa in the rat neostriatum: peaks of energy metabolism reveal a distributed functional module.

In unanaesthetized rats, mechanical stimulation of a single vibrissa increased glucose utilization in one cortical column of the somatosensory area and in several spots in the dorsolateral neostriatum, predominantly on the side contralateral to the stimulation. Two or three peaks of glucose utilization unique to the stimulated animals were seen in cross sections throughout a 1.8 mm anteroposterior extent in the dorsolateral striatum. These observations suggest that one cortical column is functionally related to several neostriatal regions. The distributed modularity may be an important characteristic of the basal ganglia system.

Behavioural effects of ablations of the presumed 'prefrontal cortex' or the corticoid in pigeons.

This study further explored functional similarities of mammalian prefrontal cortex and its presumed equivalent in pigeons. Our results show that the performance of delayed alternation of pigeons in an Y-maze is impaired following ablations of the prefrontal equivalent together with the corticoid but not of the corticoid alone. In the same maze, discrimination between vertical and horizontal stripes was unimpaired regardless of the lesion. Our results added the following new information. (1) Corticoid is not essentially involved in mediation of delayed responding. (2) Like monkeys, pigeons take much fewer trials to learn delayed alternation in a maze than in an operant chamber. (3) Lesions of the pigeon equivalent of the prefrontal cortex impair delayed responding also in the new apparatus. (4) These lesions do not impair visual pattern discrimination. Our results do not contradict the hypothesis that the postero-dorso-lateral neostriatum in pigeons is comparable to the prefrontal cortex in mammals.

Afferents to different layers of the dorsolateral isocortex in rats.

Fluorescent somatopetal tracers were used to infiltrate, by diffusion rather than injections, the dorsolateral cortex of one hemisphere in rats. In different animals the tracers penetrated into the cortex to different depths. We found several interesting features of the commissural system: first, there were no areas without commissural neurons. At least a few labelled cell bodies were present in a single-cell layer also in "acallosal" cortical areas. Secondly, there is a considerable variety of laminar distribution patterns of labelled perikarya in different areas. Thirdly, some cortical fields, which cytoarchitecturally appear uniform, can be subdivided according to different distributions of cell bodies with commissural projections. Fourthly, when only supragranular layers were infiltrated, labelled cell bodies were present mainly in the supragranular layers of the contralateral cortex. Infiltration of the first layer alone did not label any neurons in the contralateral cortex but did label neurons in layer VIb ipsilaterally. In the subcortex, the labelled perikarya were found in the structures already known to project directly to the cortex. In rats with the tracer restricted mainly to the supragranular layers, a conspicuously reduced labelling was found in the basal forebrain and the thalamus. In the thalami of those animals, labelled neurons were found only in paralamellar nuclei. The high sensitivity of the tracer used, together with infiltration of the entire dorsolateral cortex, allows us to conclude that probably all sources of innervation of the isocortex in rats have been seen.

Parcellation of the frontal cortex of the New World monkey Callithrix jacchus by eight neurotransmitter-binding sites.

The most extensive development during primate brain evolution involves the cortex of the frontal lobe, especially its prefrontal region. The distribution of neurotransmitter receptors is unknown in this part of the cortex of New World monkeys. The respective distributions of eight different receptors for the transmitters L-glutamate (L-glu and NMDA), gamma-amino-butyric acid (GABAA), noradrenaline (alpha 1), acetylcholine (M1 and M2) and serotonin (5-HT1 and 5-HT2) were therefore studied in cortical areas of the frontal lobe of the lissencephalic New World monkey, Callithrix jacchus. The results are compared to earlier data on Old World monkeys in order to obtain insight into evolutionary trends at the level of chemical neuroanatomy. Our results indicate that the density and laminar pattern of some receptors change precisely at the cytoarchitectonic boundaries between different cortical areas, while some other receptors do not exhibit measurable changes. For example, the premotor area 6 can be distinguished from prefrontal areas by its high concentration of adrenergic alpha 1 receptors as labelled with [3H] prazosin, with only the cingulate area 24 showing higher values. In other cases, the receptor distribution changes within a cytoarchitectonically homogeneous area. Thus, area 8 can be subdivided into dorsal and ventral regions on the basis of the distribution of GABAA, muscarinic and serotonin receptors. Comparison of these results in a New World monkey with receptor distributions in other primate species reveals much larger interspecies differences in the areas of the frontal lobe than e.g. in the primary visual cortex.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantitative receptor autoradiography of eight different transmitter-binding sites in the hippocampus of the common marmoset, Callithrix jacchus.

The regional and laminar distributions of eight different transmitter-binding sites were measured in the marmoset hippocampus by means of quantitative in vitro receptor autoradiography. Receptors for 5-HT1, L-glutamate, N-methyl-D-aspartate (NMDA) and GABAA were similarly distributed. The highest concentrations of these receptors were found in the pyramidal layer of CA1 and the molecular layer of the dentate gyrus. The 5-HT2 receptors showed the highest concentrations in the oriens layer of CA2. The highest concentrations of muscarinic M1 receptors were seen in the pyramidal layer of CA1. Muscarinic M2 receptors were most densely concentrated in the pyramidal layers of CA1, CA2 and CA3. The noradrenergic alpha 1 receptors were most densely packed in the radiatum-lacunosum-molecular layer of CA2 and the molecular layer of the dentate gyrus. Statistically significant co-distributions of serotoninergic, glutamatergic and GABAergic receptors point to possible interactions between these receptor systems in the same hippocampal regions and layers. Comparisons of marmoset distribution patterns for GABAA, NMDA, L-glutamate and 5-HT1 receptors with those in human hippocampi and those of other primates showed similarities between them. Clear differences in the patterns of alpha 1, M1, M2 and 5-HT2 receptors could be seen between marmoset and human hippocampi, indicating a high degree of species specificity in a presumably "conservative" brain region. More similarities, however, could be found between marmoset and human hippocampi than between those of rat and human brains, especially in relation to 5-HT1 and GABAA receptors and L-glutamate-binding sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Improved contrast in histochemical detection of cytochrome oxidase: metallic ions protocol.

The standard current technique for demonstration of cytochrome oxidase (CyOx) provides low-contrast diaminobenzidine (DAB) polymer. In order to enhance the contrast with divalent metalic ions, we have screened a number of buffers and found that Hepes, Mops and cacodylate neither precipitate these ions nor inactivate CyOx in a concentration of 0.1 M. Staining thus obtained shows a broad range of gradations between black and white. With fresh tissue the resulting image is superior to that obtained with the brown DAB product, even if a recommended blue filter or printing on very hard paper are used. The technique is as simple as the one which is currently standard. Fixed tissue, cut either in a cryostat or vibratome, can be stained well when floating but not when mounted on slides. The stained floating tissue can be used for electron microscopy, but has no advantage over the standard method.

Synaptic reorganisation of the mediodorsal thalamic nucleus in adult rat following chronic prefrontal cortical lesions.

Chronic changes in the thalamic mediodorsal nucleus (MD) after unilateral lesions of the prefrontal cortex were studied with the aid of quantitative light, and electron microscopic immunohistochemistry. Three months after the lesions, although the size of MD ipsilateral to the lesion did not change considerably, the neuronal density was significantly reduced. Conversely, as demonstrated by quantitative electron microscopy, the density of GABA immunostained axon terminals significantly increased in the lesioned side. It is suggested, that as MD does not contain GABA cells, the reactive hyperinnervation of the MD by GABA-containing axons is of extrinsic origin. This finding is also the morphological evidence of the potential of GABA-ergic nerve cells for reactive (induced) axonal sprouting.

Projections from the visual areas to the neostriatum in rats. A re-examination.

Neostriatal afferents from the primary visual cortex in rats were studied using dextran-biotin, biocytin, and Fluoro-Gold. The area V1 was found to project only to a dorsomedial, longitudinal region of neostriatum (NS), bordering on the lateral ventricle and subcortical white matter. The preterminal fibres in the NS form fluffs which increase in number and density in the cases with larger injections. This target region is poorly stained for calbindin and yet belongs to the matrix compartment. The secondary visual areas also project to the dorsomedial NS region but they also innervate the deeper tissue in the same general region. Iontophoresis of Fluoro-Gold into the dorsomedial NS labelled some pyramidal neurones in the fifth layer of the primary visual cortex. The cortical areas that surround the visual cortical complex project to other regions of the NS: the somatosensory cortex to a dorsolateral longitudinal region and the auditory area to the medial half of the caudalmost portion of NS. Thus, major sensory cortical divisions project to non-overlapping NS regions. Since NS in monkeys and cats does not receive afferents from the primary visual cortex and in a number of other species does, we conclude that visual systems in different mammals differ with respect to their projections to NS.

Anti-inflammatory drugs suppress injury-induced NADPH-d activity in CA1 pyramidal neurones.

Inducibility of NADPH-diaphorase (NADPH-d) or nitric oxide synthase (NOS) has been demonstrated in pyramidal neurones of the hippocampus, but the mechanisms of this induction are not known. The present study aimed to assess the role of anti-inflammatory drugs in injury-induced production of NADPH-d/NOS in CA1 pyramidal neurones. We found that either a steroid, dexamethasone or a non-steroid, indomethacin, prevents induction of these enzymes. We also found that NO is not necessary for the induction. None of the three drugs used had detectable effect on the neurones which contain constitutive NADPH-d/NOS.

Retention and relearning of spatial delayed alternation in rats after ablation of the prefrontal or total non-prefrontal isocortex.

In the present study we addressed the question whether, within the isocortex, the prefrontal area of the rat is uniquely involved in mediation of delayed alternation. In one group of rats the dorsolateral isocortex, from the dorsomedial shoulder to the dorsal lip of the rhinal sulcus was removed bilaterally in a single surgical session. In these rats delayed alternation in a T-maze was significantly less impaired than in rats with one stage bilateral removal of the medial prefrontal cortex. The prefrontal cortex seems not to depend crucially on isocortical input for its medication of delayed alternation.

Dopaminergic innervation of the brain in pigeons. The presumed 'prefrontal cortex'.

In the postero-dorso-lateral neostriatum (PDLNS) of the pigeon we found a high density of tyrosine hydroxylase and perineuronal nests of fibres labelled with anti-dopamine antibody. These data confirm our earlier indirect evidence that PDLNS receives a dense dopaminergic innervation and therefore can be compared with the mammalian prefrontal cortex.

A Combined golgi and postembedding GABA and glutamate electron microscopic study of the nucleus dorsomedialis thalami of the rat.

The synaptic organisation of neurons in the nucleus medialis dorsalis (MD) was investigate by combined Golgi, and post-embedding gamma-aminobutyric acid (GABA) and/or glutamate (GLU) immunogold methods. The morphological features of the impregnated neurons in the MD (detailed by Kuroda et al. 1992a) were similar to thalamocortical relay cells present in other thalamic nuclei. GABAergic cell bodies could not be found. Distal dendritic branches of gold toned neurons established synaptic contacts with small and medium size glutamate as well as GABA-immunoreactive axonal endings. Large glutamate and GABA-positive axon terminals formed synaptic contacts with proximal dendrites. GABAergic axon terminals exhibited symmetric synaptic contacts and contained pleomorphic vesicles. Glutamate immunoreactive terminals formed asymmetric synapses and contained spheroid vesicles, although, some large glutamate immunoreactive endings contained pleomorphic vesicles. Synaptic glomerulus-like complexes could also be observed with the participation of glutamate-, and GABA-positive large axon terminals synapsing with Golgi impregnated proximal dendrites.

Cortical area in the rat that mediates visual pattern discrimination.

In Experiment I, bilateral ablations of the caudolateral cortex involving Krieg's area 36 impaired discrimination of visual patterns but not delayed alternation. In Experiment II, the same type of lesions retarded postoperative learning to discriminate embedded visual patterns. In rats from the Experiment II tracers of axonal transport gave no signs of damage of the connections of the primary visual cortex. In agreement with this, Nissl stain of the dorsal lateral geniculate nuclei showed no neuronal loss or gliosis. These results suggest that caudo-lateral cortex in rats corresponds to the inferotemporal cortex of primates.

Absence of transsynaptic transport in cerebello-thalamo-cortical path of the rat.

Attempts to visualize the cerebello-thalamo-cortical path in the rat were made with different approaches. We tried (1) double labelling with somatopetal tracing from the motor cortex and somatofugal from the cerebellar nuclei, (2) transmembrane labelling by depositing biocytin or wheat germ agglutinin (WGA) into the motor cortex or cerebellum. WGA was either iodinated with 125I or conjugated with horseradish peroxidase (HRP). The double labelling technique showed an overlap of the tracers in the same thalamic region but no evidence of transsynaptic transport in either direction was obtained. Our results indicate a difference in the organization of this system in primates and rodents, since transsynaptic labelling in the cerebellar nuclei after injections of WGA-HRP conjugate in the monkey motor cortex has been found.

NADPH-diaphorase (NOS) is induced in pyramidal neurones of hippocampal slices.

We found NADPH-diaphorase (presumably identical with nitric oxide synthase) in pyramidal neurones of the hippocampus in slices that stayed in a chamber for 30 min or longer. In some instances parallel slices showed normal membrane properties when studied electrophysiologically. In freshly made slices the pyramidal neurones were not stained. Thus, after induction of the enzyme, the hippocampal pyramidal neurones can synthesize nitric oxide which may serve as a retrograde messenger in long-term potentiation. The enzyme may also play a role in cell loss seen in slices which stayed in a chamber for 9-22 h before fixation.