Superior colliculus: single unit responses to stimulation of visual cortex in the cat.

Electrical stimulation of area 18 of the cat visual cortex produces intense excitation of neurons in the superior colliculus. Excitation is followed by a period of decreased collicular responsiveness to light stimulation. These effects are seen in both directionally selective and nonselective units. The corticocollicular projection is retinotopically organized.

Somatovisceral pathway: rapidly conducting fibers in the spinal cord.

Fibers that respond to distension of hollow viscera and to mechanical stimulation of somatic structures were found primarily near the ventromedian fissure of the upper lumbar spinal cord. These fibers could be directly excited by electrical stimulation at C(1). The average conduction velocity for these fibers was 68.6 meters per second.

A new artificial antigen of the hepatitis E virus.

An artificial antigen composed of 12 small antigenic regions derived from the ORF2 and ORF3 HEV proteins was designed. The gene encoding for this artificial antigen was assembled from synthetic oligonucleotides by a new method called Restriction Enzyme-Assisted Ligation (REAL). The diagnostic relevance of this second generation HEV mosaic protein (HEV MA-II) was demonstrated by testing this antigen against a panel of 142 well defined anti-HEV positive and anti-HEV negative serum samples. The data obtained in this study support the substantial diagnostic potential of this HEV mosaic antigen.

Comparison of hand-traced and computer-based cephalometric superimpositions.

OBJECTIVE: To determine the ability to produce comparable superimpositions using hand tracing and digital methods (Dolphin v10). In addition, if the two methods were comparable, we wanted to determine if a difference existed between the best-fit cranial base superimposition and S-N superimpositions using the digital method. METHODS AND MATERIALS: Sixty-four initial (T(1)) and final (T(2)) cephalometric film radiographs were obtained. Cranial base and regional superimpositions were completed independently for each pair of radiographs by either hand tracing and digital methods. To quantitatively evaluate the differences between the two methods, the hand and digital superimpositions were digitized to obtain x-y coordinates of routine cephalometric landmarks at T(2). Linear distance between multiple corresponding (hand and digital) T(2) cephalometric landmark locations (e.g., A point) were measured and defined as the T(2) landmark distance (T(2) LD). Additionally, 61 patient records were used to compare the digital method for best-fit cranial base superimpositions versus S-N superimpositions. A Friedman test was applied to examine for differences. RESULTS: The upper 95% confidence limit for the mean of the T(2) LD for hand and digital superimposition methods was <1 mm for all landmarks except maxillary incisor tip and apex. The upper 95% confidence interval for best-fit vs S-N was >1 mm for most landmarks. CONCLUSION: This study validates the use of superimpositions produced by Dolphin Imaging version 10 and is a necessary step forward toward widespread acceptance of digital superimpositions.

[Diagnostic advantages of the test system "DS-EIA-HBsAg-0.01" for detection of HBV surface antigen].

The new highly sensitive test system "DS-EIA-HBsAg-0.01" (Priority Certificate No. 2006129019 of August 10, 2006) in detecting hepatitis B surface antigen (HBsAg) was assessed. The sensitivity of the test was estimated using the federal standards sample HBsAg 42-28-311-06, panels' samples Boston Biomedica Inc. (West Bridgewater, Mass, USA) and ZeptoMetrix Corp. (Buffalo, NY, USA). The findings have indicated that "DS-EIA-HBsAg-0.01" is equally effective in detecting different subtypes of HBsAg during a seroconversion period earlier than alternative assays. Along with its high analytical and diagnostic sensitivity, the system shows a high diagnostic specificity.

A cellular mechanism for the bidirectional pain-modulating actions of orphanin FQ/nociceptin.

Orphanin FQ/nociceptin (OFQ/N) and its receptor share substantial structural features and cellular actions with classic opioid peptides and receptors, but have distinct pharmacological profiles and behavioral effects. Currently there is an active debate about whether OFQ/N produces hyperalgesia or analgesia. Using a well-defined brainstem pain-modulating circuit, we show that OFQ/N can cause either an apparent hyperalgesia by antagonizing mu opioid-induced analgesia or a net analgesic effect by reducing the hyperalgesia during opioid abstinence. It presumably produces these two opposite actions by inhibiting two distinct groups of neurons whose activation mediates the two effects of opioid administration. OFQ/N antagonism of the hyperalgesia may have significance for the treatment of opioid withdrawal and sensitized pain.

Contributions of the amygdala and medial prefrontal cortex to incentive cue responding.

Reward-seeking behavior is controlled by neuronal circuits that include the basolateral nucleus of amygdala (BLA), medial prefrontal cortex (mPFC), nucleus accumbens (NAc) and ventral tegmental area. Using a discriminative stimulus (DS) task in which an intermittently presented cue (DS) directs rats to make an operant response for sucrose, we previously demonstrated that dopamine receptor antagonism in the NAc reduced reinforced cue responding, whereas general inactivation of the NAc increased behavioral responding in the absence of the cue. Because they send major glutamatergic projections to the NAc, the BLA and mPFC may also contribute to reward-seeking behaviors modulated by the NAc. In this study we compare the effects of BLA and mPFC inactivation on rats' performance of a DS task. BLA inactivation by combined GABA(A) and GABA(B) agonists impaired cue responding with minimal effects on operant behavior in the absence of cues. Dorsal medial prefrontal cortex (dmPFC) inactivation also inhibited cue-evoked reward-seeking. In contrast, ventral medial prefrontal cortex (vmPFC) inactivation disinhibited responding to unrewarded cues with less influence on reinforced cue responding. These findings demonstrate that the BLA and dmPFC facilitate cue-evoked reward-seeking, whereas, in the same task the vmPFC exerts inhibitory control over unrewarded behaviors.

A locus and mechanism of action for associative morphine tolerance.

Repeated administration of an opioid in the presence of specific environmental cues can induce tolerance specific to that setting (associative tolerance). Prolonged or repeated administration of an opioid without consistent contextual pairing yields non-associative tolerance. Here we demonstrate that cholecystokinin acting at the cholecystokinin-B receptor is required for associative but not non-associative morphine tolerance. Morphine given in the morphine-associated context increased Fos-like immunoreactivity in the lateral amygdala and hippocampal area CA1. Microinjection of the cholecystokinin B antagonist L-365,260 into the amygdala blocked associative tolerance. These results indicate that cholecystokinin acting in the amygdala is necessary for associative tolerance to morphine's analgesic effect.

Opposing effects of intra-nucleus accumbens mu and kappa opioid agonists on sensory specific satiety.

Mu opioid (MOP) agonists acting in the nucleus accumbens (NAcc) robustly enhance consumption of palatable foods. In addition, the effect on consumption of palatable foods produced by MOP agonists acting in the NAcc depends on both recent flavor exposure and the availability of a choice between different-flavored foods. In contrast, kappa opioid (KOP) agonists have variable effects on feeding and KOP agonists have MOP opposing behavioral actions when microinjected at several brain sites. We previously demonstrated that NAcc MOP agonists reverse the devaluation (satiety) effect of pre-feeding for a given flavor; in fact, NAcc MOP agonists selectively increase consumption of a recently sampled food. In contrast, in the present study, we found that the selective KOP agonist U50488 injected into the NAcc of rats reduced consumption of a recently sampled flavor while increasing consumption of the flavor that was not pre-fed. Intra-NAcc U50488 did not affect overall consumption or flavor preference in the absence of pre-feeding. The present data, in conjunction with our previous findings, highlight the robust and opposing role of NAcc MOP and KOP opioid receptors in palatability-based food choice and consumption and raise the possibility that an endogenous KOP agonist acting in the NAcc contributes to the phenomenon of sensory specific satiety.

Nucleus accumbens opioid signaling conditions short-term flavor preferences.

Opioid signaling in the nucleus accumbens (NAcc) strongly modulates flavor-based food choice. To further investigate the role of opioid signaling in taste reward, we used a sensory specific satiety (SSS) paradigm to devalue specific flavors of nutritionally identical food pellets in rats. In the NAcc, infusion of a mu opioid (MOP) receptor selective agonist selectively increased consumption of a pre-fed flavor, thus reversing the SSS effect. Conversely, blockade of endogenous opioid signaling with the opioid antagonist naltrexone selectively decreased consumption of a recently consumed flavor, potentiating the SSS effect. No enhancement of consumption was observed if a delay of 3 h was imposed following the intra-NAcc MOP agonist indicating that there were no long-term changes in flavor preference. If a delay was introduced between the initial flavor exposure and the intra-NAcc MOP agonist infusion, pellet consumption was increased non-selectively (irrespective of flavor) suggesting that close temporal contiguity between flavor experience and NAcc opioid action is critical for the opioid effect on flavor preference. In contrast to opioid effects, inactivating NAcc neurons by local microinjection of muscimol (a GABAA agonist) increased consumption of both the pre-fed and non-pre-fed flavors equally. These results demonstrate that opioids released in the NAcc during consumption of palatable foods produce a selective and transient increase in preference for a recently sampled flavor.

[Impact of amino acid sequence variation of a determinant(s) on the antigenic properties of hepatitis B virus HBsAg].

Impact of amino acid sequence variation on the antigenic properties of the surface hepatitis B virus antigen HBsAg was studied. Eleven recombinant HBsAg variants of wild (adr, ayw2, adw2, adw4, aywl, adw2) and vaccine escape (adw2 T126S, adw2 Q129L, adw2 Q129R, adw2 T143K, adw2 Q145R, aywl Q145A) were obtained. All the recombinant antigens were tested on a panel of 43 monoclonal antibodies (MAb) specific to different HBsAg determinants. Amino acid sequence variation of the a-determinant of HBsAg was shown to significantly affect its immunological responsiveness and antigenic properties. Amino acid substitution in different positions or in the same position, but for various amino acids may differently affect these properties.

Nucleus accumbens opioids regulate flavor-based preferences in food consumption.

Opioid signaling in the nucleus accumbens (NAcc) regulates feeding behavior, having particularly strong effects on consumption of highly palatable foods. Since macronutrient content may contribute to palatability, it is uncertain whether opioid regulation of food consumption is based primarily on its macronutrient content or its flavor per se. In order to isolate the effect of flavor, we manipulated opioid signaling in the NAcc in rats and quantified consumption of differently flavored but nutritionally identical pellets. When pellets of either flavor were presented alone, microinjection of d-Ala(2),N,Me-Phe(4),Gly-ol(5)-enkephalin (DAMGO (a mu opioid receptor (MOP) agonist)) into the NAcc increased consumption of pellets of both flavors equally. When both flavors of pellets were presented simultaneously, however, DAMGO in the NAcc selectively increased, while naltrexone (a non-selective opioid antagonist) in the NAcc selectively decreased, consumption of the more preferred flavor. Systemic naltrexone injection had no flavor specific effects, decreasing consumption of both flavors equally. Non-selective inactivation of NAcc neurons by local microinjection of muscimol (a GABA(A) agonist) increased consumption of both the more- and less-preferred flavors equally. These results indicate that opioid signaling directly regulates a subset of NAcc neurons that can selectively enhance consumption of preferred palatable foods based exclusively on flavor cues.

Roles of alpha1- and alpha2-adrenoceptors in the nucleus raphe magnus in opioid analgesia and opioid abstinence-induced hyperalgesia.

Noradrenaline and alpha-adrenoceptors have been implicated in the modulation of pain in various behavioral conditions. Noradrenergic neurons and synaptic inputs are present in neuronal circuits critical for pain modulation, but their actions on neurons in those circuits and consequently the mechanisms underlying noradrenergic modulation of pain remain unclear. In this study, both recordings in vitro and behavioral analyses in vivo were used to examine cellular and behavioral actions mediated by alpha1- and alpha2-adrenoceptors on neurons in the nucleus raphe magnus. We found that alpha1- and alpha2-receptors were colocalized in the majority of a class of neurons (primary cells) that inhibit spinal pain transmission and are excited during opioid analgesia. Activation of the alpha1-receptor depolarized whereas alpha2-receptor activation hyperpolarized these neurons through a decrease and an increase, respectively, in potassium conductance. Blockade of the excitatory alpha1-receptor or activation of the inhibitory alpha2-receptor significantly attenuated the analgesia induced by local opioid application, suggesting that alpha1-receptor-mediated synaptic inputs in these primary cells contribute to their excitation during opioid analgesia. In the other cell class (secondary cells) that is thought to facilitate spinal nociception and is inhibited by analgesic opioids, only alpha1-receptors were present. Blocking the alpha1-receptor in these cells significantly reduced the hyperalgesia (increased pain) induced by opioid abstinence. Thus, state-dependent activation of alpha1-mediated synaptic inputs onto functionally distinct populations of medullary pain-modulating neurons contributes to opioid-induced analgesia and opioid withdrawal-induced hyperalgesia.

Nucleus accumbens dopamine release is necessary and sufficient to promote the behavioral response to reward-predictive cues.

The nucleus accumbens is part of the neural circuit that controls reward-seeking in response to reward-predictive cues. Dopamine release in the accumbens is essential for the normal functioning of this circuit. Previous studies have shown that injection of dopamine receptor antagonists into the accumbens severely impairs an animal's ability to perform operant behaviors specified by predictive cues. Furthermore, excitations and inhibitions of accumbens neurons evoked by such cues are abolished by inactivation of the ventral tegmental area, the major dopaminergic input to the accumbens. These results indicate that dopamine is necessary to elicit neural activity in the accumbens that drives the behavioral response to cues. Here we show that accumbens dopamine release is causal to the rats' reward-seeking behavioral response by demonstrating that dopamine in this structure is both necessary and sufficient to promote the appropriate behavioral response to reward-predictive cues.

Profiles of NK, NKT cell activation and cytokine production following vaccination against hepatitis B.

Human natural killer (NK) cells (CD56+ CD3-) represent crucial components of the innate immune system especially against viral infections and because their activation can modulate the outcome of the adaptive immune response. NKT cells (CD56+CD3+), a lymphocyte T population characterized by expression of surface markers of NK cells, are known to be abundant in the liver and their activation could be associated with hepatic injury. Using three-color flow cytometry to measure surface receptors and intracellular cytokines, we have explored early activation signals and cytokine production in NK and NKT cells within a group of hepatitis B vaccinated and non-vaccinated individuals. A specific increase of the CD56bright cell population, the activation receptor CD69 and IFN-gamma, was observed in NK cells following incubation with recombinant HBsAg in responders to vaccination. Comparable results were observed in NKT cells showing an increment of CD69, CD25, IL-2 and IFN-gamma expression in responder subjects. These parameters were statistically diminished in non-responder individuals (p<0.05) in both groups of cells. These results demonstrate a diminished activation of these cells in non-responders to the vaccine, suggesting that NK and NKT cells play an important role in the immune response following hepatitis B vaccination.

Axonal trajectories and terminations of on- and off-cells in the cat lower brainstem.

Two physiologically defined classes of pontomedullary raphe neurons were intracellularly labeled in order to determine the target nuclei of their axonal projections. In the lightly anesthetized cat, cells either increased (on-cells) or decreased (off-cells) their discharge rate during the paw withdrawal reflex evoked by noxious pinch or heat. On- and off-cells were injected with horseradish peroxidase and the initial course of labeled axons through the lower brainstem was reconstructed. On-cell projections to the pontomedullary raphe and medial reticular nuclei were sparse. On-cells projected densely to regions of the lateral reticular formation and the ventrolateral medulla at both rostral and caudal medullary levels. In general, on-cells had few collaterals and spare axonal swellings. In contrast to on-cells, most off-cells had axons that collateralized densely within the brainstem raphe and adjacent reticular formation. Such collaterals were either local, within the neuron's dendritic field, or distant, involving a projection of 1-8 mm. One off-cell had a dense terminal field within the sensory trigeminal complex, a projection that may subserve the inhibition of trigeminal sensory neurons produced by raphe magnus stimulation. Well-labeled off-cells had numerous collaterals and dense regions of axonal swellings. In summary, off-cells terminated densely in the raphe magnus and adjacent reticular formation whereas on-cells projected predominantly to the ventrolateral medulla, a region implicated in autonomic control. Local off-cell collaterals provide an anatomical substrate that would enable off-cells to coordinate the activity of on- and off-cells through synaptic contacts.

The origin of descending pathways in the dorsolateral funiculus of the spinal cord of the cat and rat: further studies on the anatomy of pain modulation.

There is considerable evidence that the dorsolateral funiculus (DLF) of the spinal cord contains descending pathways critical for both opiate and brainstem stimulation-produced analgesia. To obtain a comprehensive map of brainstem neurons projecting to the spinal cord via the DLF, large injections of horseradish peroxidase (HRP) were made into the lumbosacral spinal cord of cat and rat. These injections were made caudal to midthoracic lesions which spared only a single DLF or ventral quadrant (VQ); thus only those neurons whose axons descended in the spared funiculus would be labelled. Cells with descending axons in the VQ were concentrated in the medullary nucleus raphe pallidus and obscurus, nucleus retroambiguus and in various subregions of the reticular formation including the nucleus reticularis ventralis, gigantocellularis, magnocellularis, pontis caudalis and pontis oralis. Significant numbers of neurons were also found in medial and lateral vestibular nuclei and in several presumed catecholamine-containing neurons of the dorsolateral pons. In the rat, but not in the cat, considerable numbers of cells are present in the mesencephalic reticular formation just lateral to the periaqueductal gray. In both species, some cells were found in the paraventricular nucleus of the hypothalamus. Brainstem cells projecting in the DLF were concentrated in the nucleus raphe magnus and in the adjacent nucleus reticularis magnocellularis, ipsilateral to the spared funiculus. Significant numbers of cells were found in the dorsolateral pons, differing somewhat in their distribution from those projecting in the VQ. DLF-projecting cells were also present in the ipsilateral Edinger-Westphal nucleus and periaqueductal grey contralateral red nucleus of the midbrain and in the ipsilateral hypothalamus. Smaller projections from other sites are described. These results are discussed in terms of the differential contribution of several brainstem neuronal groups, including the serotonergic nucleus, raphe magnus, the ventromedial reticular formation of the medulla, and various catecholamine-containing neurons of the dorsolateral pontine tegmentum to the analgesia produced by opiates and electrical brain stimulation.

Somatodendritic morphology of on- and off-cells in the rostral ventromedial medulla.

The rostral ventromedial medulla (RVM) contains two classes of physiologically defined neurons, on-cells and off-cells, that are implicated in nociceptive modulation. In a continuing effort to detail the neural circuitry that underlies the activity of these two distinct neuronal types, the somatodendritic morphology of on- and off-cells was studied in the cat, rat, and ferret. In lightly anesthetized animals, on-cells increased and off-cells decreased their discharge rate during a withdrawal reflex evoked by noxious stimuli. Following their physiological characterization by using intracellular recording, on- and off-cells were injected with either horseradish peroxidase or biocytin and their somatodendritic arborizations were examined. Labeled on- and off-cells included fusiform and stellate cells of all sizes as well as large multipolar neurons. Although the somatic shape of both on- and off-cells in RVM was heterogeneous, off-cells tended to be fusiform neurons whose long axis was oriented mediolaterally. The dendritic domains of both on- and off-cells extended bilaterally past the lateral edge of the trapezoid body or pyramid and ventrally to, and sometimes including, the trapezoid body or pyramid. In contrast to their extensive mediolateral spread, the dendritic domains of both cell types were limited to the ventral half of the reticular formation and were compressed along the rostrocaudal axis. The dendritic arbor of individual on- and off-cells extended well beyond the cytoarchitectonic boundaries of any single nuclear region, within the domain delineated as the RVM. The spatial domains of the dendritic arbors of on- and off-cells are further evidence that the on- and off-cells throughout the RVM constitute an integrated unit in the modulation of nociceptive transmission.

GABA-immunoreactive boutons contact identified OFF and ON cells in the nucleus raphe magnus.

The pontomedullary raphe magnus (RM) contains two physiologically defined types of neurons that participate in the opioid-induced modulation of dorsal horn nociceptive messages: OFF cells, which decrease, and ON cells, which increase their discharge rates when reflex behavior is evoked by noxious pinch or heat. Because both types of neuron have inhibitory inputs and because there is evidence that gamma-aminobutyric acid (GABA) inhibitory mechanisms within RM contribute to the antinociceptive action of opioids, we have sought anatomical evidence for a direct GABAergic input to OFF and ON cells. In this study, cells of each type located in the RM were electrophysiologically defined and intracellularly filled with horseradish peroxidase or Neurobiotin. One cell of each type was labeled in the cat, and 2-3 cells of each type were labeled in the rat. Thin sections were labeled by a postembedding immunogold procedure by using an antibody directed against glutaraldehyde-conjugated GABA. GABA-immunoreactive (GABA-ir) boutons contained small, round, clear vesicles and made symmetrical synapses with identified dendrites. GABA-ir boutons were apposed to soma and to proximal and distal dendrites of both cell types in both species. These findings demonstrate direct GABAergic input to identified OFF and ON cells in the RM. J. Comp. Neurol. 378:196-204, 1997.