Olfactory neuroblastoma in dogs and cats--a histological and immunohistochemical analysis.

Olfactory neuroblastoma (ONB) was identified in 13 dogs and nine cats. The tumours were subjected to microscopical examination and were graded using a human pathological grading system. In the canine and feline tumours there was more necrosis and higher mitotic activity (mitotic index and Ki67 labelling index) than reported in human ONB. Rosettes were a common feature of feline ONBs. A significant correlation was observed between the histological grade and the Ki67 labelling index. The histopathological diagnosis of ONB was confirmed immunohistochemically by demonstration of the neuronal marker neuron-specific enolase (NSE). Two other neuron-specific antibodies specific for microtubule-associated protein-2 (MAP-2) and neuronal nuclei antigen (NeuN) were evaluated. MAP-2 expression proved to have higher specificity than labelling for NSE. NeuN expression was less sensitive and of limited practical value.

[Endoscopic diagnostics and treatment of flat intestinal lesions]. / Endoskopická diagnostika a lécba plochých nádorových lézí strevních.

The aim of the article is to provide information on current options for endoscopic diagnostics and treatment of flat tumor intestinal lesions. These neoplasms are frequently and inaccurately described as sessile polyps, which have different genetic and diagnostic features, different prognosis and therapy. Although until recently, the lesions have been managed using surgery, the current approach is predominantly endoscopic.

Granulomatous pneumonia caused by Mycobacterium genavense in a dwarf rabbit (Oryctolagus cuniculus).

A juvenile dwarf rabbit (Oryctolagus cuniculus) with clinical signs of dyspnea and suspected ascites was submitted for necropsy. The main macroscopic findings were a watery red pleural effusion and some whitish striated foci in the lungs. In addition, there were multifocal scars in the cortex of the kidneys. The histologic examination of the lungs showed a severe granulomatous pneumonia with detection of acid-fast bacilli, in the kidneys, an interstitial chronic lymphoplasmacellular nephritis with interstitial fibrosis, and in the brain, a multifocal granulomatous and partly necrotizing encephalitis with detection of spores, suggestive of encephalitozoonosis. In the lungs, Mycobacterium genavense was verified by polymerase chain reaction and 16S ribosomal RNA gene sequencing. To our knowledge, this is the first report of an M. genavense infection in a rabbit, with the lungs being the only affected organ. Therefore, an aerogen infection seems to be the most contemplable way of infection.

Exogenous corticosteroid and shifts of circadian rhythms in hamsters.

We tested the hypothesis that glucocorticoid stimulation mediates the effect of exercise on circadian clock resetting in hamsters. We injected animals with 1 and 5 mg dexamethasone--a potent glucocorticoid agonist--at zeitgeber time (ZT) 4 and ZT6, circadian phases at which vigorous exercise induces maximal phase advances of about 3 h. Neither dose of dexamethasone induced phase shifts that were significantly larger than those induced by injections of saline vehicle at either of the phases tested. Some animals, however, showed quite large and consistent phase shifts to repeated injections whether with saline or dexamethasone, such that there was a statistically significant correlation between individuals' responses to the two treatments. The data indicate no role for increased glucocorticoid activity in mediating the effects of exercise on circadian phase shifting, but suggest a modest role for nonspecific stimulation, independent of exercise, in inducing phase shifts at ZT4-ZT6.

Cellular colocalization of Fos and neuropeptide Y in the intergeniculate leaflet after nonphotic phase-shifting events.

Nonphotic and photic stimuli that phase shift circadian rhythms were presented to hamsters, Mesocricetus auratus. The nonphotic stimulus was a 3-h pulse of novelty-induced wheel running starting at circadian time 4-5. The photic stimulus used was a 0.5 h light pulse starting at circadian time 18. Double immunocytochemistry was used to determine the neurochemical phenotype of cells in the intergeniculate leaflet that were activated by these stimuli. Both the nonphotic and the photic phase-shifting stimuli induced the expression of c-fos in the intergeniculate leaflet compared to unstimulated controls. However, after nonphotic stimulation, Fos-like immunoreactivity was common in neurons that also were NPY positive. Such colocalization of Fos and NPY after photic stimuli was rare. These findings suggest that the NPY pathway from the intergeniculate leaflet to the suprachiasmatic nucleus carries information about nonphotic events.

Rabbit and ferret parietal cell inhibition by Helicobacter species.

We tested sonicates of Helicobacter pylori, H. mustelae, and H. felis for inhibition of acid secretion in rabbit and ferret isolated gastric glands. Three H. pylori strains, two of three H. mustelae strains, and two H. felis strains significantly inhibited acid secretion in rabbit cells by 95.2-93.3%, 55.9% and 96.4%, and 83.4-96%, respectively. All Helicobacter strains examined inhibited acid secretion by ferret cells by 65.3-76.8%, 89.1-97.6%, and 85.8-92.8%. H. pylori inhibited acid secretion after stimulation with histamine and isobutylmethylxanthine or with 8-bromo-cyclic adenosine monophosphate (P < 0.05 for all tests). These findings demonstrate that acid inhibition is a property common to the three Helicobacter species tested. It occurs independently of the mammalian origin of the parietal cell, and it does not involve blockade of histamine-2 receptors.

Neuropeptide Y and behaviorally induced phase shifts.

Neuropeptide Y-containing fibers project from the intergeniculate leaflet of the lateral geniculate nucleus to the suprachiasmatic nucleus. Previous studies have indicated that this pathway may be involved in non-photic resetting of the circadian clock. Therefore, we investigated the possibility that neuropeptide Y mediates phase shifts induced by a particular non-photic stimulus, a pulse of running in a novel wheel. Confining hamsters to a small nest box failed to block phase shifts induced by neuropeptide Y given at zeitgeber time 4; this indicates that increased locomotor activity is not necessary for the observed shifts. Antiserum raised against neuropeptide Y or normal serum was administered at circadian time 5 through a cannula aimed at the suprachiasmatic nucleus. The hamsters were then removed from their cages and placed in a novel wheel for 3 h. Hamsters that received normal serum and ran > 5000 revolutions in the novel wheel advanced their rhythms (mean shift 2.55 h +/- 0.22 S.E.M.) by amounts similar to those of unoperated hamsters. Administration of neuropeptide Y antiserum attenuated the shift normally associated with running in a novel wheel (mean shift 0.21 h +/- 0.14 S.E.M.). These studies indicate that the neuropeptide Y input from the lateral geniculate nucleus to the biological clock is involved in the phase shifts seen in response to novelty-induced wheel running. It also provides another example of the ability of antisera to alter behavior. This may be a useful approach in manipulations of neurochemical activity when antagonists are not yet available or poorly defined.

Intergeniculate leaflet lesions and behaviorally-induced shifts of circadian rhythms.

The aim of this work was to assess the effect of lesions of the intergeniculate leaflet on nonphotic phase shifts produced by confining hamsters to novel running wheels for 3 h in the middle of the subjective day. In intact hamsters this procedure produces large phase advances provided that the hamsters maintain high levels of wheel running during the confinement. Intergeniculate leaflet lesions blocked or reduced phase shifts after confinement to a novel wheel. However, for most animals these lesions also reduced both the amount of activity during the 3 h pulse in the novel wheel and the amount of daily wheel running in the home cage. To boost activity of lesioned hamsters to levels associated with large phase shifts, the animals were confined to novel wheels at low ambient temperature. The lesioned hamsters still failed to show large phase shifts. The benzodiazepine triazolam also failed to induce phase shifts in lesioned animals, but it induced less activity in lesioned animals as compared to sham-operated controls. The data support the hypothesis that the intergeniculate leaflet conveys information about nonphotic phase-shifting to the circadian pacemaker in the suprachiasmatic nucleus. They also raise the possibility that some effects of intergeniculate leaflet lesions previously interpreted as having a photic basis, might be due to the activity-lowering effect of the lesions.

Retinohypothalamic projections and immunocytochemical analysis of the suprachiasmatic region of the desert iguana Dipsosaurus dorsalis.

Two separate and distinct retinal projections to the hypothalamus in the iguanid lizard Dipsosaurus dorsalis were described using horseradish peroxidase and cobalt-filling techniques. Both of the projections were unilateral and completely crossed; one terminated in the supraoptic nucleus and the other in the suprachiasmatic nucleus. Immunocytochemical analysis showed that the supraoptic nucleus contained cell bodies and fibers that cross-react with antibodies raised against arginine vasopressin, while the suprachiasmatic nucleus contained arginine vasopressin-like immunoreactive fibers emanating from cells in the nearby paraventricular nucleus. The suprachiasmatic nucleus contained a dense plexus of fibers that cross-reacted with neuropeptide-Y antibody. Antiserum against vasoactive intestinal polypeptide showed no reactivity in any part of the forebrain, while antiserum against serotonin showed sparse and uniform reactivity throughout the forebrain, including the suprachiasmatic nucleus. These results, together with other data, indicate that the suprachiasmatic nucleus of D. dorsalis is homologous to the suprachiasmatic nuclei of rodents, structures known to contain circadian pacemakers. We suggest that the suprachiasmatic nucleus may play a similar role in the circadian system of D. dorsalis.

Phase angle changes of photically entrained circadian rhythms following a single nonphotic stimulus.

Syrian hamsters entrained to a light-dark (LD) cycle of 14:10 h were given the opportunity to run in novel wheels for 3 h in the middle of the light phase. This manipulation transiently altered the phase angle of entrainment to the LD cycle: activity onset was significantly advanced (by about 0.5 h) on the day after the pulse and gradually drifted back toward its prepulse time. When animals were held in LD 11.5:12.5 h, a photoperiod in which onset time occurs later relative to the time of lights-off, they again advanced about 0.5 h in response to the pulse of wheel running, but many animals retained an advanced phase angle for at least 7 days, and some for more than 21 days. Individual changes in phase angle were highly correlated with the prepulse phase angle: the more negative the phase angle, the greater the advance subsequent to the novel wheel pulse. These results show that a single, short-duration, nonphotic manipulation can produce long-lasting alterations in the phase angle of entrainment to a LD cycle.

Nonphotically induced phase shifts of circadian rhythms in the golden hamster: activity-response curves at different ambient temperatures.

Running in a novel wheel during the subjective day can shift the circadian activity rhythm of a hamster. The amount of running is thought to be an important variable. We generated a dose-response (activity-phase shift) curve for the amount of wheel running during a 3 h period starting 8 h before normal dark onset in a 14:10 LD cycle. At room temperature (23 degrees C) the relationship was sigmoidal: from 0 to 4000 revolutions resulted in minimal phase advances (up to 50 min). From 4000 to 5000 revolutions the magnitude of the advances increased sharply, and above 5000 revolutions phase advances were asymptotic at about 3 h. The same general relationship held when hamsters were stimulated to be more active in the novel wheel by lowering the ambient temperature to either 11 degrees C or 6 degrees C. However, at these lower temperatures, a significant number of animals did not shift more than the minimal amount of 50 min even though they ran more than 5000 revolutions. This indicates that running per se in a novel wheel was not sufficient to induce phase shifts. Possibly, at room temperature, the amount of wheel running reflects a particular motivational state produced by the rewarding nature of wheel running, although at low ambient temperatures at least some individuals run primarily to meet thermoregulatory needs.

Behavioral decoupling of circadian rhythms.

Golden hamsters (Mesocricetus auratus) were kept in a light-dark cycle (LD 14:10). For 2 weeks, almost every day they were placed in a novel running wheel for 3 hr, starting 7 hr before dark onset. Most of the animals made several thousand wheel revolutions during this 3 hr. When these animals were subsequently transferred to a dark room, their activity was split into two components, one close to the time of the previous exposure to the novel wheel and the other close to the time when they had been active in the dark phase of the previous LD cycle. The two components fused after a few days in darkness. These observations show that nonphotic events are capable of causing major reorganizations of circadian activity patterns, despite the presence of an LD cycle.

Gene expression in the geniculate induced by a nonphotic circadian phase shifting stimulus.

We examined the effect of a nonphotic stimulus (running in a novel wheel) on the induction of Fos-related proteins in the two well-established neural components of the circadian clock: the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) of the lateral geniculate complex. There was no induction of Fos immunoreactivity (IR) in the SCN, but the IGL showed distinct Fos IR in animals whose running levels were associated with maximal rhythm shifts. Induction of Fos immunoreactivity was greatest at circadian phases when wheel running induces phase shifts. This temporal and spatial specificity of Fos induction is evidence that the intergeniculate leaflet mediates nonphotic signals to the circadian clock.

Circadian rhythms of plasma melatonin in the ruin lizard Podarcis sicula: effects of pinealectomy.

Plasma melatonin was measured in lizards (Podarcis sicula) at six different times of day under conditions of constant temperature and darkness. Intact animals showed a circadian rhythm of melatonin with a peak in the subjective night of 207 pg/ml (median) and a trough during the subjective day that was below the minimum detection level of the assay (50 pg/ml). Pinealectomy abolished the circadian rhythm of plasma melatonin; median levels were near or below the minimum detection level at all times sampled. The data suggest that the pineal is the only source of rhythmic blood-borne melatonin in Podarcis sicula, and are consistent with the hypothesis that changes in the free-running period of the locomotor rhythm induced by pinealectomy in this species are due to withdrawal of rhythmic melatonin from the blood.

The effect of pinealectomy on circadian plasma melatonin levels in house sparrows and European starlings.

We determined 24-hr plasma melatonin profiles in intact, sham-pinealectomized, and pinealectomized European starlings (Sturnus vulgaris) and house sparrows (Passer domesticus) in a light-dark (LD) cycle and in constant darkness (DD). In the intact and sham-pinealectomized birds of both species, a melatonin rhythm was found, with low levels during the day and high levels during the night. Pinealectomy abolished the nighttime peak of melatonin in both species; hence, levels were low at all times sampled. This uniform response of plasma melatonin to pinealectomy contrasts with the differential response of circadian activity rhythms to pinealectomy for these two species. In DD, locomotor activity in pinealectomized house sparrows is usually arrhythmic, whereas in starlings a rhythm usually persists. This suggests that in the latter species free-running circadian rhythms are not necessarily dependent on a rhythm in plasma melatonin. The same is true for the synchronized activity rhythm observed in pinealectomized birds of both species in LD, as well as for the damped rhythm that persists in pinealectomized house sparrows following an LD-to-DD transfer. The results are consistent with the hypothesis that the pineal and its periodic output of melatonin constitute only one component in a system of at least two coupled pacemakers. They also suggest that there are species differences in the relative role played by the pineal and other pacemakers in controlling circadian rhythms in behavior.

Circadian locomotor rhythms in the desert iguana. II. Effects of electrolytic lesions to the hypothalamus.

Desert iguanas, Dipsosaurus dorsalis, displaying freerunning circadian locomotor rhythms in conditions of constant darkness and temperature received electrolytic lesions to the hypothalamus. The locomotor activity of those lizards (N = 9) which sustained 80% or more damage to the suprachiasmatic nucleus (SCN) became arrhythmic whereas all animals that sustained less than 35% damage to the SCN remained rhythmic, even though they sustained significant damage to nearby regions of the hypothalamus and preoptic area. These results suggest strongly that the SCN plays a role in the regulation of circadian rhythms in the desert iguana. Taken together with other evidence, they support the view that this structure is homologous to the mammalian SCN, which acts as a pacemaker in the circadian system.

Circadian locomotor rhythms in the desert iguana. I. The role of the eyes and the pineal.

The pineal and the eyes are known to be important components in the circadian system of some species of lizards; their effects may be mediated by the hormone melatonin. We examined the role played by these structures in the desert iguana (Dipsosaurus dorsalis). Surgical removal of the pineal had no effect on circadian locomotor rhythms, even though this procedure abolished the circadian rhythm of melatonin in the blood. Furthermore, when the isolated pineal of Dipsosaurus was studied in organ culture, it showed no circadian rhythm of melatonin secretion, as do pineals of some other lizard species, although it did produce large quantities of this hormone. Bilateral ocular enucleation had only small effects on the freerunning period of locomotor rhythms, without affecting melatonin levels in the blood. Behavioral circadian rhythms persisted in desert iguanas subjected to both enucleation and pinealectomy. These data suggest that neither the pineal nor the eyes are central components of the circadian pacemaking system in Dipsosaurus, nor is melatonin critically involved in maintaining its organization.

Effect of iodine disinfection products on higher plants.

Iodine is used to disinfect potable water on United States spacecraft. Iodinated potable water will likely be used to grow plants in space. Little is known about the effects of iodine disinfection products on plants. Seeds of select higher plants were germinated in water iodinated using the Shuttle Microbial Check Valve, and water to which measured amounts of iodide was added. Percent germination was decreased in seeds of most species germinated in iodinated water. Beans were most affected. Germination rates, determined from germination half-times, were decreased for beans germinated in iodinated water, and water to which iodide was added. Development was retarded and rootlets were conspicuously absent in bean and several other plant species germinated in iodinated water. Iodide alone did not elicit these responses. Clearly iodine disinfection products can affect higher plants. These effects must be carefully considered for plant experimentation and cultivation in space, and in design and testing of closed environmental life support systems.