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Interfacing electronics with neural tissue is crucial for understanding complex biological functions, but conventional bioelectronics consist of rigid electrodes fundamentally incompatible with living systems. The difference between static solid-state electronics and dynamic biological matter makes seamless integration of the two challenging. To address this incompatibility, we developed a method to dynamically create soft substrate-free conducting materials within the biological environment. We demonstrate in vivo electrode formation in zebrafish and leech models, using endogenous metabolites to trigger enzymatic polymerization of organic precursors within an injectable gel, thereby forming conducting polymer gels with long-range conductivity. This approach can be used to target specific biological substructures and is suitable for nerve stimulation, paving the way for fully integrated, in vivo–fabricated electronics within the nervous system.

Injectable bioelectronics could become an alternative or a complement to traditional drug treatments. To this end, a new self-doped p- type conducting PEDOT-S copolymer (A5) was synthesized. This copolymer formed highly water-dispersed nanoparticles and aggregated into a mixed ion-electron conducting hydrogel when injected into a tissue model. First, we synthetically repeated most of the published methods for PEDOT-S at the lab scale. Surprisingly, analysis using high-resolution matrix-assisted laser desorption ionization-mass spectroscopy showed that almost all the methods generated PEDOT-S derivatives with the same polymer lengths (i.e., oligomers, seven to eight monomers in average); thus, the polymer length cannot account for the differences in the conductivities reported earlier. The main difference, however, was that some methods generated an unintentional copolymer P(EDOT-S/EDOT-OH) that is more prone to aggregate and display higher conductivities in general than the PEDOT-S homopolymer. Based on this, we synthesized the PEDOT-S derivative A5, that displayed the highest film conductivity (33 S cm(-1)) among all PEDOT-S derivatives synthesized. Injecting A5 nanoparticles into the agarose gel cast with a physiological buffer generated a stable and highly conductive hydrogel (1-5 S cm(-1)), where no conductive structures were seen in agarose with the other PEDOT-S derivatives. Furthermore, the ion-treated A5 hydrogel remained stable and maintained initial conductivities for 7 months (the longest period tested) in pure water, and A5 mixed with Fe3O4 nanoparticles generated a magnetoconductive relay device in water. Thus, we have successfully synthesized a water-processable, syringe-injectable, and self-doped PEDOT-S polymer capable of forming a conductive hydrogel in tissue mimics, thereby paving a way for future applications within in vivo electronics. Funding Agencies|Swedish Research Council [2018-05258, 2018-06197]; Swedish Foundation for Strategic Research (e-NeuroPharmacology) [RMX18-0083]; European Research Council (ERC) project [e-NeuroPharma 834677]

Because the reactors of the Swedish reactor program were erected over a limited period of time, there are significant similarities regarding used materials and manufacturing methods between the dif ...

The combined effect of thermal aging and irradiation on cast and welded stainless steel solidification structures is not sufficiently investigated. From theory and consecutive aging and irradiation ...

The effect of dissolved hydrogen (DH) on primary water stress corrosion cracking of nickel base alloys has been of intense interest for plant operators worldwide. In this study, crack growth rates of Alloy 600 were measured in simulated PWR primary coolant at 330 °C with DH levels of 5, 16, 45 and 75 cc H2/kg H2O, respectively. The oxide films formed in the crack tip regions were examined using transmission electron microscopy (TEM). The results show low and similar crack growth rates at all DH levels, without a maximum at 16 cc H2/kg H2O. The low DH content favors nickel oxide formation at the crack tip region, whereas the high DH level favors Me3O4 type spinel formation. Also, the oxide films were found to grow epitaxially on some metal grain surfaces in the cracks. The possible effects of alloy composition on the oxide films formed, and the effect of DH on the crack growth are briefly discussed.

Friction is an important phenomenon in everyday life. All children are familiar with playground slides, which may thus be a good starting point for investigating friction. Motion on an inclined plane is a standard physics example. This paper presents an investigation of friction by a group of 11-year olds. How did they plan their investigations? What aspects of friction could they discern? What understanding of the nature of science was revealed—and developed—during their investigation and subsequent discussion with the teacher?

Crystalline lenses with multiple focal lengths in monochromatic light (multifocal lenses) are present in many vertebrate groups. These lenses compensate for chromatic aberration and create well-focused color images. Stabilization of the lens within the eye and the ability to adjust focus are further requirements for vision in high detail. We investigated the occurrence of multifocal lenses by photorefractometry and lens suspension structures by light and electron microscopy in sturgeons (Acipenseriformes, Chondrostei) as well as sharks and rays (Elasmobranchii, Chondrichthyes). Multifocal lenses were found in two more major vertebrate groups, the Chondrostei represented by Acipenseriformes and Chondrichthyes represented by Elasmobranchii. The lens suspension structures of sturgeons, sharks, and rays are more complex than described previously. The lens is suspended by many delicate suspensory fibers in association with a ventral papilla in all groups studied. The arrangements of the suspensory fibers are most similar between sturgeons and sharks. In rays, the lens is suspended by a smaller ventral papilla and the suspensory fibers are arranged more concentrically to the lens. J. Morphol., 2012. © 2012 Wiley Periodicals, Inc.

To elucidate possible mechanisms behind the endocrine control of parr-smolt transformation, the daily plasma profiles in thyroid hormones (TH; free thyroxine (FT(4)), total thyroxine (TT(4)), and total 3,5,3'-triiodothyronine (TT(3))), growth hormone (GH) and cortisol were studied in Atlantic salmon parr and smolts under simulated-natural winter (8 L:16D) and spring (16.5 L:7.5D) photoperiods, respectively. Overall, TT(4), TT(3) and GH levels were higher in smolts than in parr, whereas FT(4) levels fluctuated within the same range in parr and smolts. Significant diurnal changes in plasma TH were present in parr. Both FT(4) and TT(4) levels increased during the photophase and decreased during the scotophase, while TT(3) levels followed an inverse pattern. Growth hormone showed no significant changes in parr. Changes in FT(4), TT(4), GH, and cortisol, but not TT(3), levels, were observed in smolts with peak levels during both the photophase and scotophase for FT(4), TT(4) and GH. Plasma cortisol was not assayed in parr but in smolts the peaks were associated with dusk and dawn. In addition to the general increases in TH, GH and cortisol, the distinct endocrine differences in nighttime levels between parr in the winter and smolts in the spring suggest different interactions between TH, GH, cortisol and melatonin at these different time points. These spring scotophase endocrine profiles may represent synergistic hormone interactions that promote smolt development, similar to the synergistic endocrine interactions shown to accelerate anuran metamorphosis. The variations in these diurnal rhythms between parr and smolts may represent part of the endocrine mechanism for the translation of seasonal information during salmon smoltification.

Box jellyfish have the most elaborate sensory system and behavioural repertoire of all cnidarians. Sensory input largely comes from 24 eyes situated on four club-shaped sensory structures, the rhopalia, and behaviour includes obstacle avoidance, light shaft attractance and mating. To process the sensory input and convert it into the appropriate behaviour, the box jellyfish have a central nervous system (CNS) but this is still poorly understood. The CNS has two major components: the rhopalial nervous system and the ring nerve. The rhopalial nervous system is situated within the rhopalia in close connection with the eyes, whereas the ring nerve encircles the bell. We describe the morphology of the ring nerve of the box jellyfish Tripedalia cystophora as ascertained by normal histological techniques, immunohistochemistry and transmission electron microscopy. By light microscopy, we have estimated the number of cells in the ring nerve by counting their nuclei. In cross sections at the ultrastructural level, the ring nerve appears to have three types of neurites: (1) small "normal"-looking neurites, (2) medium-sized neurites almost completely filled by electron-lucent vacuoles and (3) giant neurites. In general, only one giant neurite is seen on each section; this type displays the most synapses. Epithelial cells divide the ring nerve into compartments, each having a tendency to contain neurites of similar morphology. The number and arrangement of the compartments vary along the length of the ring nerve.

Cubozoans differ from other cnidarians by their body architecture and nervous system structure. In the medusa stage they possess the most advanced visual system within the phylum, located in sophisticated sensory structures, rhopalia. The rhopalium is a club-shaped structure with paired pit-shaped pigment cup eyes, paired slit-shaped pigment cup eyes, and two complex camera-type eyes: one small upper lens eye and one large lower lens eye. The medusa carries four rhopalia and visual processing and locomotor rhythm generation takes place in the rhopalia. We show here a bilaterally symmetric organization of neurons, with commissures connecting the two sides, in the rhopalium of the cubozoan Tripedalia cystophora. The fortuitous observation that a subset of neurons is strongly immunoreactive for a PCNA (proliferating cell nuclear antigen)-like epitope allowed us to analyze the organization of these neurons in detail. Distinct PCNA-immunoreactive (PCNA-ir) nuclei form six bilateral pairs that are associated with the slit eyes, pit eyes, upper lens eye, and the posterior wall of the rhopalium. Three commissures connect the clusters of the two sides and all clusters in the rhopalium have connections to the area around the base of the stalk. This neuronal system provides an anatomical substrate for integration of visual signals from the different eyes.

In order to understand the development of retinal cells, we have studied the temporal expression of HuC/D protein in embryonic, postnatal and adult rat retina. During development and in the adult retina, practically all cell somata in the ganglion cell layer and the vast majority of conventional amacrine cells in the inner nuclear layer displayed HuC/D immunoreactivity. Most but not all ganglion cells expressed HuC/D at embryonic day 15, suggesting a delay between final mitosis and the initiation of HuC/D expression. Immunoreactivity for HuC/D was also evident in developing but not mature horizontal cells. Combined immunohistochemical visualization of HuC/D protein and the growth-associated protein (GAP-43) showed a distinct localization of GAP-43 in a specific compartment close to the somato-dendritic region of developing HuC/D-positive cell somata. The localization of GAP-43 immunoreactivity to a specific soma compartment became less evident during maturation. Immunoreactivity for HuC/D and GAP-43 was also discernible in horizontal cells at postnatal day 14. In the adult retina, most GAP-43 immunoreactivity was seen in the inner plexiform layer. Detailed analysis showed that HuC/D and GAP-43 expression is restricted to subsets of retinal neurons during development and in the mature retina. Thus, GAP-43 appears to be correlated with initial steps of differentiation and outgrowth of dendritic processes in HuC/D-positive ganglion and amacrine cells.

Salmonid parr-smolt transformation (smoltification) is a transitional stage between freshwater and seawater life when the salmon imprint on their natal stream, change from a territorial feeding behavior to downstream migration, increase their hypoosmoregulatory competence, and change body shape and coloration. Photoperiod is recognized as the most potent initiator and regulator of the smoltification processes. Environmental light signals giving information about photoperiod are conveyed through a light-brain-pituitary axis that encompasses the neural pathways between photoreceptor organs, the brain and the pituitary, and the diurnal rhythm in melatonin secretion by the photosensory pineal organ. In addition, changes in photoperiod may elicit changes in this axis. Employing retinal neural tract tracing and growth associated protein-43 (GAP-43) immunocytochemistry, we provide here new evidence of a structural reorganization in the light-brain-pituitary axis during smoltification. Retinal tract tracing revealed that projections in smolts expand into new territories of the ipsilateral nucleus preopticus parvocellularis pars anterior (PPa) and additional fibers invade deeper into the contralateral PPa and the nucleus preopticus magnocellularis (PM). At this time, GAP-43-immunoreactive cells appear transiently in specific cell groups throughout the brain, but mainly in the olfactory bulb, telencephalon, and hypothalamus. These structural changes in the light-brain-pituitary axis and hypophysiotropic systems are followed by sequential surges of brain neurotransmitters and receptors, and occur prior to the major surges of circulating thyroid hormone and growth hormone levels that are central to smoltification-related processes. Our data point to a specific period of structural reorganization in certain brain circuits, and we hypothesize that these are involved in triggering the subsequent behavioral, endocrine, and physiological changes associated with smoltification. (C) 2003 Published by Elsevier Science B.V.

Comparing two objects falling together is a small-scale version of Galileo’s classical experiment, demonstrating the equivalence between gravitational and inertial mass. We present here investigations by a group of ten-year-olds, who used iPads to record the drops. The movie recordings were essential in the follow-up discussions, enabling the students to compare the different situations and to discern situations where air resistance was essential and where it could be neglected. By considering a number of familiar situations and simple investigations that can be performed, e.g., on a playground,students may come closer to an appreciation of the deep significance of the non-influence of mass on motion under gravity.

Zones containing actively dividing cells (proliferation zones: PZs), in the brain of adult three-spined sticklebacks, were identified by autoradiographic detection of (3)H-thymidine and immunocytochemical detection of the thymidine analogue 5'-bromodeoxyuridine (BrdU), singly or in combination, and by immunocytochemical detection of proliferating cell nuclear antigen (PCNA) by monoclonal antibodies. The PZs are associated with boundaries between adult brain regions, as well as with defined morphofunctional subdivisions. PZs are located at the border between the telencephalon and diencephalon, and at the border between the mesencephalon and the rhombencephalon. In the midbrain, the PZ follows the dorsomedial, caudal, and ventrolateral aspects of each tectal hemisphere, extending over the caudal aspect of the torus semicircularis to the nucleus lateralis valvulae. In the hindbrain, the major PZ apparently represents the persisting embryonic secondary matrix layer of the developing cerebellum. In the forebrain, the PZs are associated with the ventricular zones of the olfactory bulbs and ventral telencephalic area ("subpallium"), dorsal telencephalic area ("pallium"), preoptic region, ventral thalamus, dorsal thalamus, epithalamus, pretectum, posterior tuberculum, and the hypothalamus. The diencephalic PZs are parcellated according to a neuromeric organisation (a synencephalic, a posterior, and an anterior parencephalic neuromere: p1, p2, and p3). The PZs of the secondary prosencephalon (telencephalon and hypothalamus) thus would belong to neuromeres p4-6, but do not form an immediately recognised serial pattern. The prosencephalic PZs correlate well with parts of embryonic migration areas as defined by Bergquist and Kallen ([1954] J. Comp. Neurol. 100:627-659), morphogenetic fields from which postmitotic neurones migrate to their final destination.

Free plasma thyroxine (FT4) levels were measured in coho salmon, Oncorhynchus kisutch, during parr-smolt transformation (smoltification) using an equilibrium dialysis system followed by a radioimmunoassay. The FT4 data were correlated to total plasma thyroxine (TT4), triiodothyronine (TT3), and growth hormone (GH). Plasma samples were taken weekly from early April to late May, when the salmon where released from the hatchery. Free thyroxine and GH levels increased gradually through smoltification. TT4 levels increased significantly in mid-April and in mid-May. TT3 levels increased in April and remained elevated until late-May after which they declined to the lowest levels. During the first increase in TT4 levels, FT4 levels remained low and TT3 levels did not increase until FT4 levels increased in late-April. In addition, after TT4 levels decreased in late May, FT4 levels remained elevated. These data show that there are differences between the plasma FT4 and TT4 profiles during smoltification. Nevertheless, regression analysis indicates that FT4 levels are highly correlated to the increases in the levels of GH (r=0.73) and TT4 (r=0.70). In addition, GH is less correlated to TT4 and TT3 (r=0.24 and r=−0.46, respectively) compared with FT4 (r=0.73), suggesting a close relationship between the increases of FT4 and GH. In addition, these data suggest that this method of measuring free plasma thyroxine may provide a new tool for studying the timing of thyroid hormone action and regulation during parr-smolt transformation in salmonids.

The inclusive 2(3He,t) reaction has been studied at 2 GeV for energy transfers up to 500 MeV and scattering angles from 0.250 up to 40. Data are well reproduced by a model based on a coupled-channel approach for describing the NN and NΔ systems. The effect of final state interaction is important in the low energy part of the spectra. In the delta region, the cross-section is very sensitive to the effects of Δ-N interaction and ΔN → NN process. The latter has also a large influence well below the pion threshold. The calculation underestimates the experimental cross-section between the quasi-elastic and the delta peaks; this is possibly due to projectile excitation or purely mesonic exchange currents.

Putative GABAergic elements in the retina of the Atlantic salmon have been identified by immunohistochemistry, utilising polyclonal antisera against γ-aminobutyric acid (GABA) and the GABA transporter GAT-1. Cell types immunoreactive (ir) for GABA comprise horizontal cells, amacrine cells, displaced amacrine cells in the ganglion cell layer, displaced amacrine cells in the inner plexiform layer (interstitial cells), and Muller cells. In addition, a GABA-immunonegative type of interstitial cell was also identified. In the inner plexiform layer, GABAir fibres were organised in sublayers that were strikingly similar to the sublayering of GAT-1ir fibres. GAT-1ir cell bodies comprise amacrine cells and displaced amacrine cells that may represent a subpopulation of the GABAir ones. In view of the very similar sublayering of GABAir and GAT-ir fibres in the IPL we suggest that a similar type of GABA transporter, that can be recognised with antibodies against rat GAT-1, is present at least in the dendrites of all GABAir amacrine cells but is not expressed in the cell bodies of all GABAir cells.

Thyroid hormones transiently increase during parr-smolt transformation in coho salmon, Oncorhynchus kisutch, and are believed to trigger morphological, physiological, behavioural, and neural changes. The effectiveness of propylthiouracil (PTU) to induce hypothyroidism in smolting coho salmon was determined by immersing coho salmon, Oncorhynchus kisutch, in 30 mg l−1 PTU from May 1, two weeks prior to the consistent annual total thyroxine (TT4) peak in mid-May, until the last sampling date. Plasma was obtained at two sampling dates from control and PTU -treated coho salmon: May 15, during the plasma TT4 peak; and May 26, after the TT4 peak. Radioimmunoassays were used to measure plasma TT4, total triiodothyronine (TT3), free thyroxine (FT4), and salmon growth hormone (sGH). The PTU -treatment inhibited the natural smoltification-related increases in plasma TT4, TT3 and GH levels compared with controls, but PTU-treatment did not affect these hormone levels when they were low. PTU -treatment increased FT4 and decreased TT3 and sGH levels in the May 26 sample. In the May 15 sample, FT4 levels were unaffected by PTU-treatment, whereas TT4 levels were decreased. These data demonstrate the ability of PTU to induce hypothyroidism in salmonids as shown by the decrease in TT4 and TT3. These data demonstrate that PTU treatment by immersion can induce hypothyroidism in salmonids as shown by: (1) the inhibition of the natural increases of TT4 and TT3; (2) the increase in FT4 levels corresponding to the lowered TT3 levels, suggesting an inhibition of thyroxine 5′-monodeiodinase activity. We also show for the first time that PTU treatment can lower plasma GH levels in salmonids. This lowering of plasma GH level is associated with the decrease in TT3 levels and the increase in FT4 levels. The PTU induced lowering in GH levels may contribute to the observed changes in FT4 and TT3, since GH is known to increase thyroxine 5′-monodeiodinase activity.

1. In the feline lower oesophageal sphincter (LOS), the distribution of the carbon monoxide (CO) producing enzymes haem oxygenase (HO)-1 and -2 was studied by immunohistochemistry and confocal microscopy, the HO activity was measured and the possible role for CO as a mediator of relaxation was investigated. 2. HO-2 immunoreactivity was abundant in nerve cell bodies of the submucosal and myenteric plexus. Approximately 50% of the HO-2-containing myenteric cell bodies were also nitric oxide synthase- and vasoactive intestinal peptide (VIP)-immunoreactive. In addition, HO-2 immunoreactivity was seen in nerve fibres, in non-neuronal cells dispersed in the smooth muscle and in arterial endothelium. HO-1 immunoreactivity was confined to non-neuronal cells in the smooth muscle, similar to those positive for HO-2. 3. Activity of HO, measured as CO production, was observed in LOS homogenates at a rate of 1.00 +/- 0.05 nmol mg-1 protein h-1. This production was inhibited by the HO inhibitor, zinc protoporphyrin-IX (ZnPP). 4. In isolated circular smooth muscle strips of LOS, developing spontaneous tone, exogenously administered CO evoked a concentration-dependent relaxation reaching a maximum of 93 +/- 3%. This relaxation was accompanied by an increase in cyclic GMP, but not cyclic AMP levels. The relaxant response was attenuated by methylene blue, but unaffected by tetrodotoxin. Repeated exposure to CO resulted in a progressive reduction of the relaxant response. 5. ZnPP caused a rightward-shift of the concentration-response curves for the relaxant responses to VIP, peptide histidine isoleucine, and pituitary adenylate cyclase activating peptide 27. 6. ZnPP and tin protoporphyrin-IX (another inhibitor of HO) did not affect nonadrenergic, noncholinergic relaxations induced by electrical field stimulation. Nor did ZnPP affect relaxations induced by 3-morpholino-sydnonimine or forskolin. 7. The present findings, showing localization of HO immunoreactivity to both neuronal and nonneuronal cells of the feline LOS, ability of LOS to produce CO and a relaxant effect of CO in circular LOS muscle, suggest a role for CO as a peripheral messenger.

The inhibitory neurotransmitter γ-aminobutyric acid (GABA) is known to exert various neurotrophic actions in the developing nervous system, but little is known about its distribution in the central nervous system during early development. We have studied the development of GABA-immunoreactive (GABAir) neurons during embryogenesis of a teleost fish, the three-spined stickleback. As early as 51 h postfertilization (PF; hatching occurs 144–168 h PF, and the first monoaminergic neurons appear around 72 h PF) GABAir neurons appear in the ventral prosencephalon caudal to the optic recess, in the ventral meencephalon, and in the spinal cord. Then, there is a gradual addition of GABAir cell groups in the rostral prosencephalon and ventral rhombencephalon (66 h PF), dorsal and caudal hypothalamus and pretecturn (72 h PF), ventral hypothalamus (78 h PF), preoptic region, thalamus, and in the meencephalon and rhombencephalon (96 h PF). GABAir axons appear in the spinal cord already at 51 h PF, and then gradually appear in the various tracts of the early axonal scaffold of pathfinding fibers, so that by 96 h PF the entire axonal scaffold contains GABAir fibers. It appears likely that GABAergic axons contribute a major population to the formation of the axonal scaffold. Moreover, in the prosencephalon GABAir neurons are arranged in clusters that may reflect a neuromerec organization with six prosencephalic neuromeres.

High spin states of 164Yb have been populated using the 152Sm(16O,4n) reaction. Analysis has been made using γγ coincidences and, for the first time in a large detector array, angular correlations. The experimental level scheme has been extended to include a total of 15 decay cascades, for some of which the spins have been measured. Extensive cranked shell model calculations have been performed to map the quasiparticle rotational bands in a window, reaching up to 1 MeV above the yrast line, in the spin-energy plane. The experimental data are compared with the calculated bands, and most of the experimental bands are interpreted in terms of quasiparticle excitations. It is also found that the number of experimentally detected bands is in good agreement with the calculated number of bands in the observed spin-energy region.

The distribution of the carbon monoxide (CO) producing enzymes haem oxygenase (HO) type 1 and 2 were studied in the feline lower oesophageal sphincter (LOS), as were HO activity and functional effects of CO. HO-2 immunoreactivity was observed in nerve cell bodies in the submucosal and myenteric plexus, nerve fibres, non-neuronal cells surrounding smooth muscle bundles, and in arterial endothelium, HO-1 immunoreactivity was confined to non-neuronal cells in the smooth muscle layer. CO production, indicating HO activity, was demonstrated in tissue homogenates. CO relaxed the LOS, and activated the cyclic GMP system. These results show that HO is present in the LOS, and suggest that CO can be generated by neuronal and non-neuronal structures and may have a role as a peripheral messenger.

The study of damping of rotational motion applying te rotational plane mapping (RPM) method is presented and discussed. The aim of this technique is to extract the distribution of the rotational transition strength from an analysis of the shape of the “central valley” of two- and three-dimensional γ-ray spectra. The method is applied to a triple γ-coincidence data set of 162,163Tm nuclei formed in 37Cl+130Te reactions. The rotational transition strength is obtained as a function of rotational frequency for selected regions of entry states, and the width is found to be rather constant and approximately equal to 80 keV. This value is significantly smaller than the value predicted theoretically for the rotational damping width Γrot. Also the ratio between the observed depth and width of both the 2D and 3D valleys does not agree with the simple model adopted in the RPM method. These discrepancies point to the presence of both a wide and a narrow component in the distribution of rotational strength as extracted by the RPM method. The analysis of simulated spectra obtained on the basis of realistic band-mixing calculations, including residual interactions, confirms this behaviour.

In order to obtain a basis for future investigations concerning the possible interactions between melatonin, GABA and benzodiazepines in the central nervous system of a teleost fish, the Atlantic salmon, we have studied the expression of immunoreactivity with a monoclonal antibody against the GABAA-receptor beta 2/beta 3-subunits (bd-17) in the salmon brain. Immunoreactivity was found in all parts of the brain, mostly as a diffuse labelling of discrete neuropil areas but in some instances as a granular perikaryal labelling. Strong neuropil labelling is located in the telencephalon, dorsal thalamus/pretectum, optic tectum, torus semicircularis, and ventrolateral tegmentum. Perikaryal labelling was observed in the stratum periventriculare of the optic tectum, torus longitudinalis, torus semicircularis, ventrolateral tegmentum, and in the granular layer of the cerebellum. The general pattern of distribution is similar to that observed in mammals, in which high receptor densities are found in the telencephalon (cerebral cortex), superior and inferior colliculi, and cerebellum. There is a good correlation with the distribution of melatonin binding sites, observed in a previous study, in areas receiving visual input such as the optic tectum, pretectum, and torus semicircularis. Moreover, a correlation was found in the inferior lobes and regions connected with them. Regions containing both bd-17-immunoreactivity and melatonin binding sites may constitute areas of functional interaction between melatonin, GABA and benzodiazepines in the central nervous system.

The distribution of putative histaminergic neurons in the brain of a teleost, the three-spined stickleback, was investigated by means of immunocytochemistry using specific antibodies against histamine (HA), and conventional microscopy as well as confocal laser scanning microscopy. Histamine-immunoreactive (HAir) neurons form discrete populations ventral to the nucleus of the posterior recess (NRP) and in the nucleus saccus vasculosus (NSV), which belong to the periventricular hypothalamic nuclei. The neuronal somata are subependymally located, and do not possess apical neurites contacting the cerebrospinal fluid. They give rise to both long-range and local axonal projections. The local projections give rise to a field of dense punctate immunoreaction dorsal to the NRP and lateral to the NSV. Long-range projections are comprised of ascending projections to the thalamus, habenula, preoptic area and dorsal telencephalon; and descending projections via the posterior tuberal nucleus, ventrally to the nucleus interpeduncularis, and dorsally into the central gray. HAir neurons occur together with serotoninergic cerebrospinal fluid-contacting (CSFc) neurons in the NRP, and with tyrosine hydroxylase-immunoreactive (THir) neurons in the NSV. Although HAir elements occur together with THir ones in many brain areas, direct contacts between the two neurotransmitter systems are rare. The putative histaminergic neurons in the brain of the three-spined stickleback constitute a very discrete neuronal system, with a major projection area in the dorsal telencephalon in a region which is considered homologous with the dorsal pallium of land vertebrates.

The neuroanatomical organization of neurons projecting to the pituitary and the origin of pituitary dopamine and nonapeptides were investigated in the brain of the Atlantic salmon (Salmo salar). Carbocyanine tract tracing in combination with tyrosine hydroxylase, arginine vasotocin and isotocin immunocytochemistry for double labelling revealed a previously unknown organization of hypophysiotrophic cell groups and their extrahypothalamic projections, and provide the first direct identification in a teleost fish of the origin of the dopaminergic and nonapeptidergic innervation of the pituitary. The present data include identification of (1) hypophysiotrophic neurons in the ventral telencephalon and in the periventricular preoptic nucleus, (2) large (magnocellular) vasotocinergic hypophysiotrophic neurons in the most rostral extension of the preoptic area, (3) a distinct neuronal group located in a supraoptic/suprachiasmatic position in the anterior periventricular nucleus, that seems to be the major source of dopaminergic innervation of the pituitary, (4) the nonapeptidergic hypophysiotrophic neurons in the preoptic nucleus, (5) hypophysiotrophic neurons in the ventral and posterior hypothalamus of which some are of liquor-contacting type, (6) projections from hypophysiotrophic and non-hypophysiotrophic neurons in the preoptic nucleus to extrahypothalamic areas such as thalamic and periventricular pretectal nuclei, and (7) subdivisions within the preoptic nucleus that exhibit different combinations of hypophysiotrophic and extrahypothalamic efferent connections. Together with previous studies of retinohypothalamic projections and neurochemical organization of hypothalamic/preoptic areas, the present data suggest that the preoptic nucleus and the anterior periventricular nucleus in teleosts possess functional subdivisions with features that resemble those of the paraventricular, supraoptic and suprachiasmatic nuclei of other vertebrates. In the Atlantic salmon, specific dopaminergic and nonapeptidergic neuronal subdivisions are proposed to play a role for photoperiod control of endocrine activity.

The effect of benzodiazepines (BZP) on melatonin release was investigated in the pineal gland of the rainbow trout, Oncorhynchus mykiss, maintained under in vitro perifusion culture conditions. Melatonin and the methoxyindoles 5-methoxytryptophol (5-MTOL), 5-methoxyindoleacetic acid (5-MIAA), and 5-methoxytryptamine (5-MT) were determined directly in samples of the superfusion medium by HPLC with electrochemical detection. Melatonin release was significantly increased by addition of diazepam and clonazepam in a dose-related and reversible manner. The effects of benzodiazepines were more pronounced in light-adapted pineal organs, when melatonin secretion is low, than under scotopic conditions. When the perifusion medium was replaced by a medium containing low calcium, high magnesium concentrations, melatonin release was considerably decreased by 70% in light-adapted and 20% in dark-adapted pineal organs. Addition of diazepam to low Ca2+, high Mg(2+)-medium reversed the decrease of melatonin release and produced a clear rise in its secretion rate. Addition of the BZP antagonist flumazenil to the perifusion medium slightly decreased melatonin release in the light- and dark-adapted state, whereas the peripheral receptor antagonist PK 11195 did not alter melatonin release. The effect of diazepam is reduced by simultaneous addition of flumazenil to the superfusion medium, suggesting that the effects of diazepam are receptor-mediated. The methoxyindoles 5-MTOL, 5-MIAA, and 5-MT showed no significant changes of their release pattern after diazepam application in light- and dark-adapted pineal organs. These results suggest that BZP can influence melatonin production and release by an intrapineal action possibly on the melatonin synthesizing photoreceptor cell.

To identify the neural components in the brain of the Atlantic salmon ( Salmo salar ) that receive and relay photoperiodic cues, the neuroanatomical and neurochemical properties of the visual and hypophysiotrophic system have been investigated by means of neural tract tracing in combination with immunocytochemical localization of tyrosine hydroxylase, arginine vasotocin and isotocin. The systems were characterized in parr and were then studied during parr-smolt transformation. The data show that: (1) different putative dopaminergic diencephalic optic nuclei receive both retinal and pineal innervation and possess projections to the optic tectum, (2) a dopaminergic neuronal population in the lateral habenular nucleus seems to receive retinal, tectal and pineal inputs, (3) an extensive retino-hypothalamic innervation coincides with putative dopaminergic and peptidergic neurons, (4) several of the peptidergic neurosecretory neurons innervate the pituitary but possess also projections to central optic nuclei and the optic tectum, (5) a distinct group of putative dopaminergic neurons in the ventral pre-optic area receives extensive retinal innervation and possesses major projections to the pituitary, (6) during parr-smolt trans-formation, additional pinealogugal termination fields appear that coincide with retinal termination fields in the dopaminergic and peptidergic neuronal portion of the hypothalamic optic nucleus. The data suggest that dopaminergic and peptidergic neurons may serve as interfaces between the visual system, the pituitary and extra-hypothalamic brain areas. The discovery of additional pineal projections to the hypophysiotrophic portion of the retinorecipient hypothalamic optic nucleus in smolts further emphasizes that the hypothalamic optic nucleus constitutes a photo-neuroendocrine control center that may play an important role during parr-smolt transformation.

This study describes for the first time the presence and distribution of the nitric oxide (NO) synthesizing enzyme, NO synthase (NOS), in the retina of a teleost. NADPH diaphorase (NADPHd) histochemistry and NOS immunohistochemistry revealed both NOS immunoreactive and NADPHd positive structures in photoreceptor outer segments, amacrine cells, horizontal cells and ganglion cells. Since NO is known to stimulate the synthesis of cGMP, our results implicate an important role for NO in retinal function, especially in cGMP related events in the photoreceptors.

Low concentrations of the structural protein collagen have recently been reported in dinosaur fossils based primarily on mass spectrometric analyses of whole bone extracts. However, direct spectroscopic characterization of isolated fibrous bone tissues, a crucial test of hypotheses of biomolecular preservation over deep time, has not been performed. Here, we demonstrate that endogenous proteinaceous molecules are retained in a humerus from a Late Cretaceous mosasaur (an extinct giant marine lizard). In situ immunofluorescence of demineralized bone extracts shows reactivity to antibodies raised against type I collagen, and amino acid analyses of soluble proteins extracted from the bone exhibit a composition indicative of structural proteins or their breakdown products. These data are corroborated by synchrotron radiation-based infrared microspectroscopic studies demonstrating that amino acid containing matter is located in bone matrix fibrils that express imprints of the characteristic 67 nm D-periodicity typical of collagen. Moreover, the fibrils differ significantly in spectral signature from those of potential modern bacterial contaminants, such as biofilms and collagen-like proteins. Thus, the preservation of primary soft tissues and biomolecules is not limited to large-sized bones buried in fluvial sandstone environments, but also occurs in relatively small-sized skeletal elements deposited in marine sediments.

The Swedish reactor fleet consists of 7 BWRs and 3 PWRs which have been operating for up to 38 years and all have plans for long term operation (LTO).

Double-differential cross sections of the C-12(n, p) reaction have been measured at 98 MeV in the angular range 0-degrees-33-degrees for excitation energies up to 35 MeV. The spectra have been analysed in terms of multipole components, using peak fitting and decomposition techniques. The data have been compared with cross sections obtained in the distorted-wave Born approximation, using matrix elements from shell-model calculations. The unit cross section and the related strength of the spin-isospin part of the nucleon-nucleon interaction were determined from the extracted Gamow-Teller strength. Cross sections and fractions of strength exhausted by dipole and higher-multipole transitions were extracted and compared with data available in the literature.

Cnidarians are often used as model animals in studies of eye and photopigment evolution. Most cnidarians display photosensitivity at some point in their lifecycle ranging from extraocular photoreception to image formation in camera-type eyes. The available information strongly suggests that some cnidarians even possess multiple photosystems. The evidence is strongest within Cubomedusae where all known species posses 24 eyes of four morphological types. Physiological experiments show that each cubomedusan eye type likely constitutes a separate photosystem controlling separate visually guided behaviors. Further, the visual system of cubomedusae also includes extraocular photoreception. The evidence is supported by immunocytochemical and molecular data indicating multiple photopigments in cubomedusae as well as in other cnidarians. Taken together, available data suggest that multiple photosystems had evolved already in early eumetazoans and that their original level of organization was discrete sets of special-purpose eyes and/or photosensory cells.

Box jellyfish (Cubomedusae) possess a unique visual system comprising 24 eyes of four morphological types. Moreover, box jellyfish display several visually guided behaviours, including obstacle avoidance and light-shaft attractance. It is largely unknown what kind of visual information box jellyfish use for carrying out these behaviours. Brightness contrast is almost certainly involved, but it is also possible that box jellyfish extract colour information from their surroundings. The possible presence of colour vision in box jellyfish has previously been investigated using behavioural, electrophysiological and immunohistochemical methods. However, the results from these studies are to some degree conflicting and inconclusive. Here, we present results from an investigation into the visual system of the box jellyfish Chiropsella bronzie , using microspectrophotometry and immunohistochemistry. Our results strongly indicate that only one type of visual pigment is present in the upper and lower lens eyes with a peak absorbance of approximately 510 nm. Additionally, the visual pigment appears to undergo bleaching, similar to that of vertebrate visual pigments.

Cnidarians are often used as model animals in studies of eye and photopigment evolution. Most cnidarians display photosensitivity at some point in their lifecycle ranging from extraocular photoreception to image formation in camera-type eyes. The available information strongly suggests that some cnidarians even possess multiple photosystems. The evidence is strongest within Cubomedusae where all known species posses 24 eyes of four morphological types. Physiological experiments show that each cubomedusan eye type likely constitutes a separate photosystem controlling separate visually guided behaviors. Further, the visual system of cubomedusae also includes extraocular photoreception. The evidence is supported by immunocytochemical and molecular data indicating multiple photopigments in cubomedusae as well as in other cnidarians. Taken together, available data suggest that multiple photosystems had evolved already in early eumetazoans and that their original level of organization was discrete sets of special-purpose eyes and/or photosensory cells.

The sharpness and thus information content of the retinal image in the eye depends on the optical quality of the lens and its accurate positioning in the eye. Multifocal lenses create well-focused color images and are present in the eyes of all vertebrate groups studied to date (mammals, reptiles including birds, amphibians, and ray-finned fishes) and occur even in lampreys, i.e., the most basal vertebrates with well-developed eyes. Results from photoretinoscopy obtained in this study indicate that the Dipnoi (lungfishes), i.e., the closest piscine relatives to tetrapods, also possess multifocal lenses. Suspension of the lens is complex and sophisticated in teleosts (bony fishes) and tetrapods. We studied lens suspension using light and electron microscopy in one species of lamprey (Lampetra fluviatilis) and two species of African lungfish (Protopterus aethiopicus aethiopicus and Protopterus annectens annectens). A fibrous and highly transparent membrane suspends the lens in both of these phylogenetically widely separated vertebrate groups. The membrane attaches to the lens approximately along the lens equator, from where it extends to the ora retinalis. The material forming the membrane is similar in ultrastructure to microfibrils in the zonule fibers of tetrapods. The membrane, possibly in conjunction with the cornea, iris, and vitreous body, seems suitable for keeping the lens in the correct position for well-focused imaging. Suspension of the lens by a multitude of zonule fibers in tetrapods may have evolved from a suspensory membrane similar to that in extant African lungfishes, a structure that seems to have appeared first in the lamprey-like ancestors of allextant vertebrates.

We have examined the occurrence of acetylcholinesterase (AChE)-positive cells in the pineal organ of different developmental stages of the Pacific coho salmon. Large numbers of AChE cells were present in freshwater living alevins, in all stages of presmolts (n = 307-544), and in adult spawners (n = 696-1774), whereas seawater-living postmolts displayed a total lack of labeled cells. The AChE-reactive cells were evenly distributed within the pineal end-vesicle and stalk of the presmolts and adults. However, the AChE-positive cells that occurred in the pineal stalk were of a smaller type and more uniform in shape than the cells of the pineal end-vesicle. The dense populations of AChE-stained cells in the alevins were all situated in the caudal part of the pineal end-vesicle. We conclude that changes in pineal metabolism occur in postsmolt salmon that live in salt-water. It is not clear whether the observed change in pineal AChE expression is an "unspecific" change caused by the life in the sea, reflecting alterations that are related to aspects of osmoregulation, and/or is involved in the visual function of the pineal organ resulting from changes in the environmental lighting conditions, e.g., photoperiod, light-intensity, or spectral composition. This study adds to our previous findings of changes that occur in the central nervous system of the salmon during the time of the parr-smolt transformation and migration between limnic and marine environments, and indicates a possible central role of the pineal organ in the control of these events.

In coho salmon (Oncorhynchus kisutch), smolt transformation has been shown to be associated with sequential surges of neurotransmitters in the brain. In order to determine if the surge of serotonin (5-HT) is correlated with structural changes, we have used immunocytochemistry to observe changes in the serotonin immunoreactivity before, during and after the 5-HT surge. The following stages were studied: 12-month-old freshwater presmolts, 17-month-old freshwater presmolts, 18-month-old saltwater smolts, 19-month-old saltwater postsmolt, 24-month-old postsmolt, and adult spawners. In the 17-month-old samples, but not at any other stage, we found a set of transient (serotonin-immunoreactive) 5-HT-immunoreactive neurons in the lateral preoptic area, as well as a discrete population of 5-HT-immunoreactive neurons in the lateral part of the dorsal right habenular nucleus. In addition, a higher density of serotonergic fibers was found in the telencephalon at this stage compared to the following two stages. Since the transient 5-HT-immunoreactive structures presented here do not appear simultaneously with the 5-HT total brain concentration surge, we conclude that they are unlikely to be the source of the 5-HT surge, but are probably related to other developmental changes in the brain associated with smolt transformation.

A procedure for measuring γ-γ correlations with a 20-detector Compton-suppressed array of Ge detectors is described. As an example some data from the 152 Sm( 16 O, 4n) 164 Yb reaction are presented.

The photosensory pineal organ of teleost fish shows a circadian rhythm in melatonin synthesis, and melatonin is known to influence a number of physiological functions. However, the target sites for melatonin are not known. We have investigated the distribution of melatonin binding sites in the brain of the salmon, Salmo salar. Brains were collected for receptor binding assay and autoradiography at each of three time points: just after lights on, just before lights off, and in the dark at midnight (photoperiod light-dark 12:12, lights on at 08.00 h, lights off at 20.00 h). Specific binding of 2-[125I]iodomelatonin was observed in several brain areas. High densities were associated with (1) the optic tectum, (2) the preoptic area, (3) an area encompassing the magnocellular superficial pretectal nucleus (‘nucleus rotundus’) and the glomerular complex, (4) the inferior lobes of the hypothalamus, (5) the lateral mesencephalic tegmentum including the torus semicircularis, and (6) the molecular layer of the cerebellum. No binding was observed in the pineal organ or in the pituitary. We observed no differences in labeling between brains collected at different time points, except in the preoptic area where binding was high at 20.00 and 24.00 h, but low at 08.00 h, and in the corpus cerebelli, where labeling in the molecular laver was higher at 24.00 and 08.00 h than at 20.00 h. Saturation experiments with crude brain membranes indicated the presence of a single binding site with no significant differences related to the time of day, with Kd values ranging from 30 to 54 pM, and Bmax values from 7.0 to 10.8 fmol/mg protein. Nonspecific binding, determined with 0.1 µM melatonin, was d and Bmax values are in the same range as those reported for the so-called high-affinity binding site in mammalian neural tissue.

The strongly shape driving πh9/2[541]l/2− configuration with α=+1/2 exhibits some anomalous, and so far unexplained, features concerning the crossing frequency, ħωc, the aligned angular momentum, ix, and interaction strength, at the alignment of the first pair of i13/2 quasineutrons in several odd-Z rare earth-nuclei. The h9/2[541]1/2− and h11/2[523]7/2− bands have been studied in the stably deformed rare-earth nucleus163Tm to investigate these features. A difference in band crossing frequency of ∼ 80 keV between the two bands is found. Rotational bands built on these two configurations have been found to cross in the spin range I=25/2–29/2 ħ. Theγ-decay pattern between the two bands is established in the crossing region and analysed in terms of a moderate shape difference between them. A theoretical estimate of the size of the interaction strength between the two bands is presented and compared to the experimental value. The observed band structure in163Tm is very similar to that of167Lu which has 2 protons and 2 neutrons in addition. This observation is discussed in relation to the similarity of the yrast bands of the two even-even “core” nuclei162Er and166Yb, for which theγ-transition energies are identical within ∼0.2 keV below the vi13/2 crossing.

The presence of galanin‐like substances and their relation to substance P‐, vasotocin‐, and isotocin‐immunoreactive neurons and fibers in the brain of teleosts was investigated with immunohistochemical methods. Two specific antisera against synthetic porcine galanin (GAL) revealed cell bodies and fibers in the brain of four different teleost species (Salmo salar, Carassius carassius, Gasterosteus aculeatus, and Anguilla anguilla). In all four species the main location of galanin immunoreactivity was in the hypothalamo‐pituitary region. A detailed study of the distribution of galanin immunoreactivity in S. salar showed that galanin immunoreactive (GALir) perikarya were present in the nucleus preopticus periventricularis, an area that may be compared to the supraoptic nucleus in mammals, and in the nucleus lateralis tuberis, a nucleus involved in pituitary control in fishes that may be compared with the arcuate nucleus in mammals, GALir perikarya were found also in the nucleus recessus lateralis and in the nucleus recessus posterior. Numerous GALir fibers were present in the telencephalon and diencephalon, whereas only small numbers of fibers were found in the brainstem. In contrast to the situation in mammals, no GALir perikarya were observed in the brainstem areas corresponding to the noradrenergic locus coeruleus and serotonergic raphe nuclei in S. salar. We did not find any coexistence of GALir substances with arginine vasotocin or isotocin in neurosecretory neurons, as has been shown for galanin with the mammalian counterparts vasopressin and oxytocin. Also, the galanin‐like substance(s) and their structurally closest related peptide family, the tachykinins, belong to separate neuronal systems in teleosts. The presence of GALir neurons in brain areas known to be involved in pituitary control, and a massive GALir innervation of the pituitary, strongly indicate a role for galanin‐like substances in pituitary control also in teleosts. Furthermore, the presence of extrahypothalamic GALir fibers suggests involvement of galanin‐like substances in other brain functions in teleosts. In conclusion, there are general similarities between teleosts and mammals concerning the distribution of galanin‐like substances. However, there seem to be substantial differences in their distribution relative to functionally related peptides within the hypothalamo‐pituitary system. Whereas galanin appears to be colocalized and released together with vasopressin and oxytocin in mammals, in teleosts the homologous substances are contained within different sets of neurons that innervate the same target, the pituitary. (Less)

Immunocytochemistry using antibodies against Met-enkephalin and Leu-enkephalin has demonstrated a group of large enkephalin-immunoreactive neurons in the nucleus of the rostral mesencephalic tegmentum (mRMT) of two teleost fish. Salmo gairdneri and Salmo salar. Injections of cobalt-lysine in the medial optic tectum retrogradely labeled the above group of tegmental neurons. Tegmental neurons were labeled only ipsilaterally to the injection site. This indicates that enkephalinergic neurons in the nRMT project to the optic tectum, and that at least some of the enkephalinergic axons observed in the optic tectum belong to a tegmen-to-tectal pathway. Comparable enkephalinergic pathways have been described in reptiles and birds, where pretectal-mesencephalic nuclei contribute to the enkephalin-containing fibers that project to the optic tectum.

A program for two-dimensional analysis of γ-γ coincidence matrices is described. The program is tailored to the Digital Equipment VT340 colour graphic terminal and written in VAX Fortran. The program provides viewing of the matrix as well as projections in both energy coordinates, determination of centroids and volumes of peaks, full two-dimensional Gauss fits to a maximum of five peaks in an arbitrarily shaped area of the matrix, etc.

A facility for studies of neutron-induced reactions has been built at the upgraded Gustaf Werner cyclotron of the The Svedberg Laboratory, Uppsala, Sweden. Well-collimated, monoenergetic neutron beams are produced with a fairly long distance between the neutron source and the reaction target, to reduce background radiation. A magnetic spectrometer with large angular and momentum acceptance has been constructed to allow measurements of energy and angular distributions of light ions produced in neutron-induced reactions. Initially, studies will be undertaken of isovector monopole and Gamow-Teller resonances, induced by the (n,p) reaction in various nuclei. The performance is illustrated in a measurement of the doubly differential cross section of the 12C(n,p)12B reaction at 100 MeV.