Diamond layers grown by chemical vapor deposition on NbN systems and NbN/SiO2-based devices.

Deposits of individual diamond grains and continuous polycrystalline diamond layers have been generated by means of a HFCVD technique onto different types of untreated or seeded NbN surfaces. To test the feasibility of using diamond layers as protective coatings for aerospace applications, we carried out diamond deposition onto the lithographically defined NbN microelectrodes of a NbN/SiO2 multifinger device. The morphological and structural features of the diamond deposits and of the substrates were characterized by FE-SEM, XRD and Raman spectroscopy. The preferential growth of diamond on the superconductive NbN enables the selective coating of the NbN microstripes sputtered on the insulating SiO2. Moreover the diamond coating procedure is able to preserve the structural integrity of the substrate material and to retain the shaped architecture of the device. For the polycrystalline diamond layers grown on NbN a residual stress of -9.8 GPa, largely due to thermal stress, has been estimated by Raman analysis. The diamond coatings of the NbN-based architectures result to be mechanically stable.

Low temperature conductivity of carbon nanotube aggregates.

We compare, over wide temperature ranges, the transport properties of single-wall carbon nanotubes arranged in the form of aligned arrays or in the form of fibres. The experimental data show that both the forms of aggregates present a crossover in the transport mechanism from three-dimensional hopping of the electrons between localized states at high temperature to fluctuation-induced tunnelling across potential barriers at low temperature. The role of the junctions formed between the bundles in the array and between the nanotubes inside the fibres is discussed on the basis of the experimental results.

An innovative and viable route for the realization of ultra-thin supercapacitors electrodes assembled with carbon nanotubes.

Electrochemical Double Layer Capacitors (EDLC), also known as supercapacitors, have been fabricated using Single Walled Carbon Nanotubes (SWCNTs) as active material for electrode assembling. In particular a new way of fabrication of ultra-thin electrodes (< or = 25 microm) directly formed on the separator has been proposed, and a prototype of EDLC has been realized and tested. For such devices the specific capacitance is in the range 40-45 F/g and the internal resistances in the range 6-8 omega x cm2, at current density of 2 mA x cm-2.

Elemental mapping and microimaging by x-ray capillary optics.

Recently, many experiments have highlighted the advantage of using polycapillary optics for x-ray fluorescence studies. We have developed a special confocal scheme for micro x-ray fluorescence measurements that enables us to obtain not only elemental mapping of the sample but also simultaneously its own x-ray imaging. We have designed the prototype of a compact x-ray spectrometer characterized by a spatial resolution of less than 100 microm for fluorescence and less than 10 microm for imaging. A couple of polycapillary lenses in a confocal configuration together with a silicon drift detector allow elemental studies of extended samples (approximately 3 mm) to be performed, while a CCD camera makes it possible to record an image of the same samples with 6 microm spatial resolution, which is limited only by the pixel size of the camera. By inserting a compound refractive lens between the sample and the CCD camera, we hope to develop an x-ray microscope for more enlarged images of the samples under test.

Immunohistochemical localization of anabolic and catabolic enzymes for anandamide and other putative endovanilloids in the hippocampus and cerebellar cortex of the mouse brain.

An increasing body of evidence indicates that: 1) the endocannabinoid anandamide (AEA) and other unsaturated N-acylethanolamines (NAEs), 2) 12-(S)-lipoxygenase (12-LOX) products of arachidonic acid, and 3) unsaturated N-acyldopamines (NADAs), act as endogenous ligands of transient receptor potential vanilloid type 1 (TRPV1) channels at intracellular binding sites. AEA is synthesized and released "on demand" in neurons from its membrane precursor, N-arachidonoyl-phosphatidylethanolamine, by an N-acyl-phosphatidylethanolamine-specific phospholipase D (NAPE-PLD), and is inactivated by intracellular hydrolysis by fatty acid amide hydrolase (FAAH), whereas catechol-O-methyl-transferase (COMT) was suggested to inactivate NADAs. However, it is not known whether these enzymes or 12-LOX co-localize to any extent with TRPV1 receptors in the brain. In this study we used immunohistochemical techniques (single peroxidase and double immunofluorescence staining), and analyzed the localization of the TRPV1 channel in mouse hippocampal and cerebellar neurons with respect to NAPE-PLD, FAAH, 12-LOX and COMT. Cycloxygenase-2 (COX-2), another putative AEA-degrading enzyme, was also studied. Co-localization between TRPV1 and either NAPE-PLD or FAAH, COX-2, 12-LOX and COMT was found in Ammon's horn (CA3) hippocampal pyramidal neurons and (with the exception of 12-LOX) in some Purkinje cells. At the cellular level, both anabolic and catabolic enzymes appeared as fine grains with immunoperoxidase labeling and were observed in the somatodendritic compartment of CA3 pyramidal cells as well as (with the exception of 12-LOX) in the cytoplasm of Purkinje neurons, in which FAAH and COX-2 immunoreactivities were, however, preferentially localized in the large extension of the dendritic arbor. Our data agree with the hypothesis that, in potential "endovanillergic" neurons, endogenous TRPV1 agonists, and AEA in particular, act as intracellular mediators by being produced from and/or degraded by the same mouse brain cells that express TRPV1 receptors.

Polycrystalline diamond on self-assembled detonation nanodiamond: a viable route for fabrication of all-diamond preformed microcomponents.

Surface assisted self-assembly of detonation nanodiamond particles (with typical sizes in the range 4-10 nm) has been obtained using different fractions of colloidal aqueous dispersions as starting material. The relationship between dispersion properties and structure/geometry of the aggregates deposited on Si or glass plates has been investigated. A series of differently shaped free-standing nanodiamond structures has been prepared, analysed and used as templates for the growth of polycrystalline diamond layers by the chemical vapour deposition (CVD) technique. The possibility of obtaining textured coating with a relatively strong [Formula: see text] preferred orientation (within a solid angle of about 0.6 srad) is also reported. Overall, the coupling of nanodiamond self-assembling to the CVD diamond growth enables one to produce specimens with complex 3D architectures. The proposed microfabrication methodology could represent a viable route for the production of free-standing all-diamond microcomponents, with tailored shapes and predefined crystalline features, to be used for advanced electronic applications.

Arvanil, anandamide and N-arachidonoyl-dopamine (NADA) inhibit emesis through cannabinoid CB1 and vanilloid TRPV1 receptors in the ferret.

Cannabinoid (CB) agonists suppress nausea and vomiting (emesis). Similarly, transient receptor potential vanilloid-1 (TRPV1) receptor agonists are anti-emetic. Arvanil, N-(3-methoxy-4-hydroxy-benzyl)-arachidonamide, is a synthetic 'hybrid' agonist of CB1 and TRPV1 receptors. Anandamide and N-arachidonoyl-dopamine (NADA) are endogenous agonists at both these receptors. We investigated if arvanil, NADA and anandamide were anti-emetic in the ferret and their mechanism of action. All compounds reduced the episodes of emesis in response to morphine 6 glucuronide. These effects were attenuated by AM251, a CB1 antagonist that was pro-emetic per se, and TRPV1 antagonists iodoresiniferatoxin and AMG 9810, which were without pro-emetic effects. Similar sensitivity to arvanil and NADA was found for prodromal signs of emesis. We analysed the distribution of TRPV1 receptors in the ferret brainstem and, for comparison, the co-localization of CB1 and TRPV1 receptors in the mouse brainstem. TRPV1 immunoreactivity was largely restricted to the nucleus of the solitary tract of the ferret, with faint labeling in the dorsal motor nucleus of the vagus and sparse distribution in the area postrema. A similar distribution of TRPV1, and its extensive co-localization with CB1, was observed in the mouse. Our findings suggest that CB1 and TRPV1 receptors in the brainstem play a major role in the control of emesis by agonists of these two receptors. While there appears to be an endogenous 'tone' of CB1 receptors inhibiting emesis, this does not seem to be the case for TRPV1 receptors, indicating that endogenously released endocannabinoids/endovanilloids inhibit emesis preferentially via CB1 receptors.

Immunohistochemical localization of cannabinoid type 1 and vanilloid transient receptor potential vanilloid type 1 receptors in the mouse brain.

Cannabinoid type 1 receptors and transient receptor potential vanilloid type 1 channels have been proposed to act as metabotropic and ionotropic receptors, respectively, for two classes of endogenous polyunsaturated fatty acid amides, the acylethanolamides and the acyldopamides. Furthermore, we and others have shown that functional crosstalk occurs between these two receptors when they are expressed in the same cell. Although demonstrated in sensory neurons of the dorsal root ganglia, spinal cord and myenteric neurons, co-expression of cannabinoid type 1 and transient receptor potential vanilloid type 1 has not yet been studied in the brain. In the present study, we addressed this issue by using commercially available specific antibodies whose specificity was confirmed by data obtained with brains from cannabinoid type 1(-/-) and transient receptor potential vanilloid type 1(-/-) mice. Double cannabinoid type 1/transient receptor potential vanilloid type 1 immunofluorescence and single cannabinoid type 1 or transient receptor potential vanilloid type 1 avidin-biotin complex immunohistochemistry techniques were performed and both methods used point to the same results. Cannabinoid type 1/transient receptor potential vanilloid type 1 expression was observed in the hippocampus, basal ganglia, thalamus, hypothalamus, cerebral peduncle, pontine nuclei, periaqueductal gray matter, cerebellar cortex and dentate cerebellar nucleus. In particular, in the hippocampus, cannabinoid type 1/transient receptor potential vanilloid type 1 expression was detected on cell bodies of many pyramidal neurons throughout the CA1-CA3 subfields and in the molecular layer of dentate gyrus. In the cerebellar cortex, expression of cannabinoid type 1/transient receptor potential vanilloid type 1 receptors was found surrounding soma and axons of the vast majority of Purkinje cell bodies, whose cytoplasm was found unstained for both receptors. Cannabinoid type 1 and transient receptor potential vanilloid type 1 immunoreactivity was also detected in: a) the globus pallidus and substantia nigra, in which some intensely transient receptor potential vanilloid type 1 immunopositive cell bodies were found in dense and fine cannabinoid type 1/transient receptor potential vanilloid type 1 positive and cannabinoid type 1 positive nerve fiber meshworks, respectively; b) the cytoplasm of thalamic and hypothalamic neurons; and c) some neurons of the ventral periaqueductal gray. These data support the hypothesis of a functional relationship between the two receptor types in the CNS.

Selective nicotinamide adenine dinucleotide phosphate-diaphorase histochemical labeling of Müller radial processes and photoreceptors in the earliest stages of retinal development in the tadpole.

To investigate potential sources of nitric oxide production in the early stages of retinal development we used, in the tadpole, nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry that was reported to reveal nitric oxide synthase isoforms in the retina. In the first stages of optic vesicle differentiation, prior to optic cup invagination, histochemical positivity was detected in the radial processes of Müller cells, that provide a scaffold for migrating retinal neuroblasts, and was soon followed by intense staining of photoreceptors. These events preceded retinal laminar patterning and the appearance of histochemical positivity in other retinal cell populations. The findings indicate that nitric oxide synthase is expressed during early retinogenesis at selective sites, which are implicated in the guidance of migrating cells and in phototransduction.

Nitric oxide synthase activity reveals an asymmetrical organization of the frog habenulae during development: A histochemical and cytoarchitectonic study from tadpoles to the mature Rana esculenta, with notes on the pineal complex.

In the adult frog, structural asymmetry of the left dorsal habenula in respect to the right counterpart has been repeatedly documented in previous studies. In the present investigation, histochemical expression of beta-nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase activity was examined in the habenulae of the developing and adult Rana esculenta. In tadpoles and during metamorphosis, selective neuropil staining was consistently found within a lateral compartment of the medial subnucleus of the left dorsal habenula. The staining was still present in the same location, but much less intense, in the mature frog, indicating that the neurochemical pattern observed during development was at least in part transient. Thus, the present data point out a peculiar neurochemical pattern of the habenular asymmetry in the frog, suggesting that nitric oxide may be involved in the developmental shaping which leads to an asymmetrical configuration of the habenulae. In addition, NADPH-diaphorase-positive cells were detected in the frontal organ (the extracranial component of the pineal complex in strict relationship with the habenulae in the frog), and labeled fibers were found in the frontal nerve, which arises from the frontal organ. This latter finding supports the postulated relationship of the habenular asymmetry with the occurrence of the frontal organ. The finding of NADPH-diaphorase histochemical reactivity confined to a distinct portion of the medial subnucleus of the left dorsal habenula prompted a reexamination of the cytoarchitecture of the developing and mature habenular complex in the frog. The bicompartmentalization detected with histochemistry in the medial subnucleus of the left dorsal habenula of the developing and adult frog was fully supported by the study of Nissl-stained epithalamic sections. These data point out that the left-right structural differences of the frog habenular complex are more complex than previously believed, and may be subserved by chemically regulated developmental processes.

Seasonal variations in the frontal organ of the frog: structural evidence and physiological correlates.

Histological study of the frontal organ in the frog, Rana esculenta, was performed during spring, summer, autumn and winter. In semithin sections stained with toluidine blue, cells containing a vacuole were clearly detected during spring, and considerably increased during summer. Such cellular elements were absent in the frontal organ during autumn and winter. This morphological evidence of seasonal variation was supported by extracellular recording in the frontal organ in different seasons. Spontaneous firing rate was found to increase from the spring to the summer, and to decrease from the autumn to the winter. Altogether, these data indicate that the frontal organ may represent an autonomic component of the pineal complex with a secretory function producing neurohormonal messages involved in the annual mechanism of the reproduction.

Diencephalic asymmetries.

Structural asymmetry in diencephalic regions has been reported in a number of studies since the pioneering observations by Kemali and Braitenberg, Atlas of the frog's brain. Springer Verlag: 1969. Anatomical differences between the left and right habenulae have been identified in many lower vertebrate species. While there are few reports of structural asymmetry in the dorsal thalamus, there is evidence that asymmetrical thalamofugal projections can be induced in the visual system of chicks by lateralized sensory stimulation prior to hatching. Finally, there have been consistent reports of differences between and right sides of the hypothalamus in their sensitivity to the effects of circulating gonadal hormones in rats. In most cases, these asymmetries are sex-linked and correspond to a lateralization of function. Although the significance of these diencephalic asymmetries is still enigmatic, their existence indicates that asymmetry is not a phylogenetically recent feature of the brain, and the left-right differences in the brain may be mediated by a common ontogenetic mechanism and may underlie the development of highly specialized functions.

A new cutaneous nerve fiber connection with the frontal nerve in the frog Rana esculenta: a morphological study.

The frontal nerve arises from the frontal organ, which represents the extracranial component of the pineal complex in some lower vertebrates, and interconnects the frontal organ with the epiphysis and the brain. The existence of a previously unreported nerve branch of the frontal nerve is described here in the frog Rana esculenta and called the lateral nerve. The course of the lateral nerve and its junction with the frontal nerve have been consistently detected by means of different techniques: toluidine blue staining in semithin sections, the Landau-Ignesti method for myelinated nerve fibers, the methylene blue intravital staining for peripheral nerves, and in vitro tracing with the carbocyanine Dil. A method to preserve intact the delicate lateral nerve during dissection is also described. The lateral nerve was consistently found to be unilateral, and to join the frontal nerve at one end (either on the left or the right side), whereas the other extremity was found to be dispersed in the dermis. Thus, the lateral nerve could represent a new pathway interconnecting the skin and the brain and/or the frontal organ in the frog.

The asymmetry of the habenular nuclei of female and male frogs in spring and in winter.

Male and female frog brains stained according to the Nissl method and cut transversely, 15 microns thick, at the level of the habenular nuclei, were investigated in spring and winter. Right and left habenular nuclei were examined. The volume and the standard deviations were calculated in each portion of the habenular nuclei investigated. The frog habenula consists of a single cell group: one on the right and two on the left side of the brain which differ, among themselves, both in the volumes of the neuropil and of the cellular ring. Functional corollaries of this striking asymmetry are still unknown. However, female and male frogs' habenular nuclei are longer and larger in spring--when frogs are sexually active--than in winter. We propose that structural brain asymmetries may be sex linked and may be induced by steroid hormonal effect in the central nervous system.

A horseradish peroxidase study of the olfactory system of the frog, Rana esculenta.

The olfactory system of the frog Rana esculenta was studied by using horseradish peroxidase (HRP) tracing of axonal pathways. Injections of HRP were made in the main olfactory bulb (MOB), accessory olfactory bulb (AOB), anterior olfactory nucleus (AON), the amygdala (AMY), and in a zone of the leteral wall of the telencephalic hemisphere immediately posterior to the AOB. Projections from these sites are described and are generally similar to those obtained by degeneration methods. However, HRP reveals more extensive olfactory connections than previously reported. Ipsilateral, contralateral, and bilateral projections are described. The MOB, AOB, and AON have ipsilateral connections to each other. The MOB and AOB have very different projections. The MOB and AON project via the habenular commissure (HC) to the contralateral medial wall of the telencephalon. Ipsilateral MOB fibers also terminate in this cell-free zone where the medial forebrain bundle (MFB) originates. The AOB projects to the lateral cortex of the contralateral telencephalic hemisphere via the HC and also to the ipsilateral AMY and lateral forebrain bundle (LFB) from where some fibers project contralaterally. HRP injections in the AMY retrogradely fill cells in the ipsilateral AOB, two nuclei of the ipsilateral hypothalamus and a nucleus of cells caudal to the ipsilateral nucleus isthmi. Fibers are also labeled that project to the contralateral AMY. Few fibers were observed to decussate in the interpeduncular nucleus or optic chiasma. No olfactory fibers were found to project to the habenular nuclei, and no labeled neurons were found to project to the olfactory bulbs. No morphological asymmetry was observed qualitatively in the distribution of olfactory fibers in the two halves of the brain.

Filter for microliter volumes.

The construction of a filter for 1-30 microliters volumes of liquid is described in detail. It is a modification of the readily available Millipore Swinex filter made by adding parts constructed from PE20 Intramedic tubing which are formed in a heated die. The die can be constructed easily in the laboratory.

The distribution of substance P in the habenulo-interpeduncular system of the frog shown by an immunohistochemical method.

The distribution of substance P(SP)-like immunoreactivity was studied in the habenulo-interpeduncular (HAB-ITP) complex of the frog Rana esculenta by means of the immunohistochemical peroxidase-antiperoxidase (PAP) method. The HAB of the frog were subdivided into dorsal and ventral nuclei, corresponding to the medial and lateral HAB of mammals respectively. In addition, the dorsal HAB of the frog were morphologically asymmetric, since only the left has a lateral and a medial subnucleus. SP-like immunoreactive cells were found in the right dorsal HAB, in the lateral subnucleus of the left dorsal HAB and in a nucleus of scattered cells located rostrodorsally to the ITP. However, the medial subnucleus of the left dorsal HAB and a portion of the right dorsal HAB facing the 3rd ventricle, did not show SP immunoreactivity. Immunoreactive fibers were found in the medial subnucleus of the left dorsal HAB, in the two fasciculi retroflexes and in the ITP. Moreover, immunoreactive terminals were observed in the ventral HAB of both sides, in the ITP and on the blood vessels which cross the ITP. It is assumed that a portion of fibers connecting the various structures of the HAB-ITP complex belongs to the SP peptidergic system as suggested for mammals. However, in contrast to the mammals, the results on the frog are in favour of an histochemical asymmetry of the SP-like immunoreactivity, since a different distribution of this peptide was found between the right and left side of the brain. Such histochemical asymmetry was less pronounced than the morphological asymmetry and its functional meaning is unknown.

Quantitative evaluation of the number of fibers in the right and left fasciculus retroflexus using horseradish peroxidase.

A quantitative evaluation of the number of fibers in the right and left fasciculus retroflexus (FR) was conducted in the frog Rana esculenta after injections of horseradish peroxidase (HRP) in the habenular nuclei of the two sides of the brain and in the interpeduncular area in separate animals. It seems that the known morphological asymmetry of the habenular nuclei is not accompanied by a corresponding asymmetry of the FR and that there is an equivalent number of fibers in the FR after HRP injections in either the habenulae or the interpeduncular area.

Neuroanatomical identification of the frog habenular connections using peroxidase (HRP).

A study of the habenular nuclei connections by means of horseradish peroxidase (HRP) has never been carried out in amphibia. In the present paper we have investigated the afferent projections of the left and right habenular nuclei of the frog Rana esculenta using this technique. Cells, labelled by HRP, were either in a Golgi-like pattern or in a granular pattern. It appears that the habenular nuclei on the two sides of the epithalamus do not show different connections even though they are morphologically asymmetric. In fact, each habenula is connected bilaterally with the septal area and the bed nucleus of the hippocampal commissure, and ipsilaterally with the hypothalamic areas, the entopeduncular nucleus, the periventricular gray of the third ventricle and the interpeduncular nucleus. However, the habenular commissure has typical commissural fibres which apparently do not involve the medial portion of the left habenula. The habenular connections in the frog are generally similar to those reported in the literature for mammals. In addition, our results show the possibility that HRP is transported both retrograde and anterograde.