Responses of lateral hypothalamic glucose-sensitive and glucose-insensitive neurons to chemical stimuli in behaving rhesus monkeys.

1. Extracellular single neuron activity was recorded in the lateral hypothalamic area (LHA) of awake, behaving monkeys, with particular regard to the feeding-related functional characteristics of glucose-sensitive (GS) versus glucose-insensitive (GIS) neurons. Firing rate changes were recorded by means of carbon fiber, multibarreled glass microelectrodes during 1) microelectrophoretic application of various chemicals, 2) gustatory and olfactory stimulation, and 3) a high fixed-ratio schedule (FR) bar press feeding task. 2. In 336 neurons examined, 91 (27%) were suppressed by electrophoretically administered glucose, and so they were designated as GS cells. The 245 neurons (73%) in which the firing rates did not change during glucose applications were pronounced GIS. The 179 GS and GIS cells tested exhibited different responses to the catecholamines (CAs), noradrenaline (NA) and dopamine (DA), both of which are intimately involved in the control of feeding. More GS neurons responded to NA than did GIS cells; the predominant effect of both CAs on GS neurons was inhibition. 3. The taste responsiveness of 111 LHA neurons was examined. Fifty-seven cells (52%) showed responses to gustatory stimulation. Of 50 GS neurons tested, 33 (66%) exhibited firing rate changes to tastes. On the contrary, only 24 (39%) of the 61 GIS neurons examined responded to gustatory stimuli. Activity changes of GS neurons commonly occurred to two or more tastants, in distinction to the relative gustatory specificity shown by GIS cells. 4. Two hundred fifty-six (84%) of the 303 neurons tested responded during one or more phases of the bar press feeding task. Most activity changes occurred during the bar press (BP) and reward (RW) periods, however numerous phasic responses to cue light (CL) and cue tone (CT) were also observed. A higher proportion of the GS neurons showed task-related activity changes than did the GIS cells (77, 95% and 179, 81%, respectively). GS neurons responded more during the BP phase and to the food reward; GIS cells were more responsive during the CL that enabled acquisition and the CT that signaled reward. Thus GS neurons were responsive during the acquisition and consumption of reward, whereas GIS cells responded to external cues signaling both of these events. The gustatory neurons displayed specific task-related activity changes only in the CL (GIS cells) and BP phases (GS neurons), that is, in phases most intimately involved in sensory-motor integration. 5. Two-thirds of the 30 GS neurons tested were responsive to both gustatory and olfactory stimulation as opposed to only one-third of GIS cells.(ABSTRACT TRUNCATED AT 400 WORDS)     

Wall effects in sucrose density gradient centrifugation of viruses

Summary The depletion of sedimenting particles to be expected from wall effects in the course of sucrose gradient centrifugation, was computed for various types of swing-out rotors on the assumption that particles hitting the tube wall are lost from the gradient.This assumption was roughly verified for heat-denatured, but not for native poliovirus. Tween 20 reduced the depletion, and prevented tight sticking of heat-denatured virus particles to tube walls.     

Extracellular taurine increase in rat hippocampus evoked by specific glutamate receptor activation is related to the excitatory potency of glutamate agonists

Taurine increases in brain extracellular space due to glutamate agonists were studied in vivo in the rat hippocampus using a dialysis technique, both in the absence and in the presence of glutamate receptor antagonists. Extracellular taurine levels increased during perfusions of agonists, listed in descending order of potency: kainate (KA), N-methyl-D-aspartate (NMDA), and quisqualate (QA). While taurine increases due to KA or QA perfusions were inhibited by 6,7-dinitro-quinoxaline-2,3-dione (DNQX), those induced by NMDA were abolished in the presence of 3-(carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP). These results indicate that increases in extracellular taurine levels evoked by NMDA, KA or QA in the rat hippocampus are caused by activation of their specific receptors. Field potentials, concomitantly recorded, were quickly abolished during NMDA or KA perfusions (0.1 mM), while QA (0.25 mM) induced the appearance of bicuculline-like evoked responses. Since taurine has been proposed as an osmoregulatory substance in the rat brain, and cell swelling is known to be an early component of glutamate agonists neurotoxicity, the increases in extracellular taurine reported here could be due to taurine released through an osmoregulatory process, counteracting the neurotoxic cellular oedema induced by glutamate agonists.     

The vigilance-promoting drug modafinil counteracts the reduction of tyrosine hydroxylase immunoreactivity and of dopamine stores in nigrostriatal dopamine neurons in the male rat after a partial transection of the dopamine pathway

We studied the ability of the vigilance-promoting drug modafinil to modulate the anterograde and retrograde changes in tyrosine hydroxylase (TH) immunoreactivity and in dopamine (DA) stores in the nigro-neostriatal DA neurons, following a partial hemitransection of this ascending DA system, using a combined morphometrical, biochemical and behavioural analysis. Modafinil was given daily i.p. in doses of 10–100 mg/kg, starting 15 min after the lesion, and the partially hemitransected rats were killed 2 weeks later. Changes in TH-immunoreactive nerve cell bodies and nerve terminals induced by the partial hemitransection were studied in the substantia nigra and neostriatum in combination with image analysis. The substantia nigra and neostriatum were also subjected to biochemical analysis of DA, 3,4-dihydroxyphenylacetic acid and homovanillic acid levels. Modafinil treatment dose-dependently (10–100 mg/kg) counteracted the hemitransection-induced disappearance of nigral TH-immunoreactive nerve cell body profiles and neostriatal TH-immunoreactive nerve terminal profiles. A 2-week treatment with 100 mg/kg of modafinil also counteracted the hemitransection-induced depletion of DA stores in the neostriatum and the ventral midbrain. Moreover, the repeated daily treatment with modafinil (100 mg/kg) protected against the hemitransection-induced disappearance of striatal 5-hydroxytryptamine, 5-hydroxyindoleacetic acid and noradrenaline levels. Striatal DA function was analysed by studying apomorphine-induced (1 mg/kg, s.c.) ipsilateral rotational behaviour 4 and 11 days after the operation. A marked dose-dependent reduction of ipsilateral rotational behaviour was demonstrated after the daily modafinil treatment in the partially hemitransected rats. In another model involving unilateral nigral microinjections of 6-hydroxydopamine, acute (one single dose) modafinil (100 mg/kg) did not affect the contralateral rotational behaviour induced by apomorphine (0.05 mg/kg s.c.), when given 30 min before the apomorphine. Taken together, morphological, neurochemical and behavioural evidence has been obtained that anterograde and retrograde changes induced in the DA stores and TH immunoreactivity of the nigro-neostriatal DA neurons by a partial hemitransection are counteracted by modafinil in a dose dependent way with 100 mg/kg producing a significant protective action against impairment of DA transmission. The results of this study open up the possibility that modafinil may protect against the anterograde and retrograde degeneration of nigrostriatal DA neurons seen after mechanically induced injury.     

Mixed endocrine tumors

Mixed endocrine tumors are tumors composed of at least two distinct tumor populations, one of which is endocrine. Because of their rarity and unusual presentation, endocrine mixed tumors raise many problems of diagnosis, management and therapy. Three main types of endocrine mixed tumors are recognized: The existence of these various types has been confirmed by recent molecular studies, even if the same studies have also shown that the histogenesis of a mixed endocrine tumor cannot be predicted from its histological features. Composite tumors are the less rare mixed tumors. The recent WHO classification recommends to restrict the term of composite endocrine tumor to the epithelial tumors containing at least 30% of obviously tumoral endocrine cells; some authors recommend to use higher thresholds, of at least 50%, in order to avoid overdiagnosis. The endocrine component is usually well differentiated, easily identified by its suggestive histological features; the endocrine nature of tumor cells is confirmed by the immunodetection of specific endocrine and neuro-endocrine markers (such as chromogranin A and synaptophysin). In some cases, the endocrine component is poorly differentiated: the demonstration of neuro-endocrine markers is necessary to confirm the diagnosis. Mixed tumors can occur in every anatomical site; they are more frequent in organs containing endocrine cells in the normal state (especially the digestive tract and the pancreas), but they can also be observed in organs devoid of endocrine cells (such as the mammary gland). The management of mixed endocrine tumors must take into account the more aggressive component. Mixed tumors containing a well differentiated endocrine component and an adenocarcinomatous component are to be treated like adenocarcinomas. Mixed tumors containing a poorly differentiated endocrine component must be considered as poorly differentiated endocrine carcinomas.     

The experimental receptivity ofHelicella (Helicella) itala andCepaea nemoralis (Mollusca,Helicidae) to larvae ofMuellerius sp. andNeostrongylus linearis (Nematoda,Protostrongylidae) from chamois (Rupicapra rupicapra)

Two batches ofHelicella (H.) itala (adult specimens) and two ofCepaea nemoralis (adult and young specimens) were experimentally infected with larvae I (L-I) ofMuellerius sp. andNeostrongylus linearis obtained from the lungs and faeces ofRupicapra rupicapra. In assess larval development, the number and percentage of the total number of larvae (L-I+L-II+L-III) per mollusc were studied, together with the number and percentage of L-III per snail and the days on which the different larval stages were reached. The development ofMuellerius sp. andN. linearis was greater in larvae from faeces. For both species of molluscs, the values for the percentages of the total number of larvae and L-III were higher inN. linearis than inMuellerius sp., but there were no notable differences in the days on which the various larval stages were reached. Both nematodes achieved a greater degree of development in young specimens ofC. nemoralis than in adults. Whether the larvae came from facces or the lungs,H. (H.) itala was a better intermediate host thanC. nemoralis forMuellerius sp. andN. linearis.     

Selection in blood cells from female carriers of the fragile X syndrome: inverse correlation between age and proportion of active X chromosomes carrying the full mutation.

We have studied the patterns of mutation and X inactivation in female carriers of a fragile X mutation, to try to correlate them with various phenotypic features. We used a simple assay, which shows simultaneously the size of the mutation, its methylation status, and DNA fragments that represent the normal active and inactive X chromosomes. We have observed an age dependent process, whereby the 'full' fragile X mutation is found preferentially on the inactive X in leucocytes in adult females, but not in younger ones. This phenomenon was not observed in female carriers of a 'premutation', who have little phenotypic expression. Preliminary data suggest that young females who show preferential presence of a full mutation on the active X in leucocytes may be at increased risk for mental retardation. We have also obtained preliminary evidence for an age dependent decrease in the somatic heterogeneity of full mutations, possibly owing to selection for smaller mutated fragments. If confirmed, the latter phenomenon might account for the known decrease with age of the expression of the fragile site. Our observations suggest that a gene whose expression is affected by the presence of a full mutation (possibly the FMR-1 gene) has a cell autonomous function in leucocytes, leading to a slowly progressive selection for cells where the mutation is on the inactive X chromosome.     

Separation of temperature sensitive and temperature insensitive components of the postsynaptic potentials in the frog motoneurons

Intracellular measurements were made in the in situ spinal cord of the frog at temperatures below 5 degrees C. Responses to volleys in the sciatic nerve, in the descending fibres and in the motor axons were studied. About 30% of the motoneurons responded to sciatic volleys with 1-3 ms segmental latency, which was short enough to assume electrotonic mediation of these responses. Another group of motoneurons responded with 6-8 ms latency, i.e. with the expected delay at chemical synapses at low temperature. Latency distribution of the sciatic-evoked postsynaptic potentials was clearly bimodal in contrast with that found at higher temperatures. Postsynaptic discharges occurred with rather long latency and they were attributed to chemically-mediated excitation. Some of the postsynaptic potentials to descending volleys also occurred with quite short latency, indicating possible electrotonic transmission from supraspinal centres to motoneurons. Latency distribution of the action potentials evoked from the motor axons was bimodal, corresponding to the different, i.e. antidromic and recurrent facilitatory, mechanism of these spikes. Calculated Q10 ratios for the sciatic-evoked reflex discharges and the afferent fibre volleys were about 2.3 and 1.8, respectively. We concluded that cooling helps to separate postsynaptic potentials according to their electrotonic and chemical mediation and that electrotonic excitation does not seem to have a primary role in the generation of postsynaptic discharges initiated by dorsal root volleys in the frog.     

Involvement of the catecholaminergic system in glucagon-induced thermogenesis in Muscovy ducklings (Cairina moschata)

Physiological studies have shown that glucagon is a potential mediator of nonshivering thermogenesis (NST) in birds. The present work was undertaken in order to investigate whether the observed thermogenesis results from a direct action of glucagon on avian thermoregulatory mechanisms or in fact requires the participation of other agents such as catecholamines. Our experiments were performed using cold-acclimated (CA) ducklings which developed muscle NST. A comparison was made with thermoneutral (TN) ducklings of the same age. Our principal results showed that: (1) at ambient temperature (25 degrees C), circulating norepinephrine (NE) was markedly decreased in CA ducklings (-42%), while circulating epinephrine (E) did not undergo any consistent change; (2) in CA and TN ducklings, an intraperitoneal injection of glucagon (360 microg x kg(-1)) was followed after 10 min by prominent lipolysis and a large increase in circulating NE (4- to 6-fold) and E (14- to 17-fold), which was sustained for at least 1 h. The elevation of circulating NE was less pronounced in CA ducklings. The thermogenic action of glucagon in birds is probably indirect and involves at least the mobilization of lipids and sympatho-adrenal stimulation. The changes in peripheral noradrenergic activity during cold acclimation could be associated with adaptive changes leading to NST.     

Developing an expert system for immunophenotypical diagnosis in immunodeficiency. Age-related reference values of peripheral blood lymphocyte subpopulations in Hungary

In the process of developing a decision support system based on flow cytometric data for the diagnosis of immunodeficiency, assessment of lymphocyte subpopulations in human peripheral blood provides the key for further analysis. Samples from 273 'healthy' Hungarian subjects were measured between 1998 and 1999. Immunophenotypic data are compared here (unadjusted for gender) by different age groups: I 0-6 years (n=45); II 7-18 years (n=71); III 19-35 years (n=72); IV 36-55 years (n=48); and V 56-99 years (n=37). Two-color flow cytometric analysis was performed using the Becton Dickinson Simultest IMK Plus kit (CD45/CD14, isotype control, CD3/CD19, CD4/CD8, CD3/HLA-DR, CD3/CD16+56). All lymphocyte subpopulations were measured in all blood samples identically. The quality criteria involved at least 95% of total lymphocytes in the analysis gate, homogenous CD45+ lymphocyte population (minimum of 2000 events in the gate, CD45+ >95%). The frequency of B lymphocytes was the highest, significantly, in the youngest Hungarian subjects, but there were no significant changes with age comparing the data of other II-V age groups. On the other hand, some T subpopulations changed with aging; both CD4 and CD8 subsets varied over time including the elevation of the fraction of activated T cells as well as LGL-NK cells. Some of these changes were significant by statistical tests. Interpretation of flow cytometric data is time-consuming and requires human knowledge of an expert laboratory staff. To facilitate the diagnosis of immunodeficiency, a pilot study aiming at the development of a diagnostic algorithm has been initiated. Algorithm nodes compare the frequency of each lymphocyte subpopulations to the generated reference values. This knowledge-based system describes a short summary report as a result of the comparison, and points to some values requiring further human examination to reach a final conclusion. These reference values and the expert system appear to be a recommended basis for comparing and combining results from different laboratories.