The distribution of serotonergic nerves in microencephalic rats treated prenatally with methylazoxymethanol.

Prenatal exposure of pregnant rats to methylazoxymethanol acetate (MAM) induces microencephaly in the offspring. In the present study of these microencephalic rats (MAM rats) we used quantitative autoradiography to investigate [3H] paroxetine binding sites, which are a selective marker of serotonin (5-HT) transporters (5-HTT). The binding in the accumbens, cortex, hippocampus, and dorsolateral thalamus was significantly increased in MAM rats, compared to the control rats, while there was a significant decrease in the dorsal raphe nucleus of the MAM rats. The levels of 5-HTT mRNA in the dorsal raphe nuclei were analyzed by in situ hybridization, which revealed a significant decrease in 5-HTT mRNA-positive neurons in the MAM rats compared to the control rats. The results imply serotonergic hyperinnervation in the cerebral hemispheres of MAM rats, while a target-dependent secondary degeneration of 5-HT neurons might be induced in the dorsal raphe nuclei of MAM rats.

Dopamine D3 receptor binding by D3 agonist 7-OH-DPAT (7-hydroxy-dipropylaminotetralin) and antipsychotic drugs measured ex vivo by quantitative autoradiography.

Because the dopamine D3 receptor is primarily expressed in regions of the limbic system of brain, it was proposed that it may represent a target for antipsychotic drugs that is free of extrapyramidal side effects. An ex vivo receptor binding technique employing [3H]7-OH-DPAT was used to evaluate in vivo occupancy of dopamine D3 receptors in the rat nucleus accumbens by selective D3 agonist 7-OH-DPAT (7-hydroxy-dipropylaminotetralin) and various antipsychotic drugs. With an ID50 value of 0.07 mg/kg, the selective D3 agonist (+)-7-OH-DPAT had the most potent inhibitory effect on ex vivo binding of [3H]7-OH-DPAT among all drugs tested. Clinical doses of phenothiazine drugs, such as chlorpromazine and levomepromazine, induce binding to D3 receptors in vivo, while atypical antipsychotic drugs, such as clozapine, pimozide, and sulpiride, are very weak in inhibiting ex vivo binding of [3H]7-OH-DPAT, indicating that the role of D3 receptors as targets of antipsychotic drugs free of extrapyramidal side effects may not be important.

Eosinophil activation in the tissue: synthetic steroid, budesonide, effectively inhibits the survival of eosinophils isolated from peripheral blood but not nasal polyp tissues.

We investigated the effect of a potent synthetic steroid, budesonide (BUD), on the survival of nasal polyp (NP) derived eosinophils (EOS). BUD, at the highest dose used, 10(-6) M, decreased this survival but only by approximately one third. We speculated that the relatively small inhibitory effect of budesonide on the survival of NP-EOS could be the result of these EOS being exposed to substantial amounts of GM-CSF, IL-5 or IL-3. In this regard, we detected 148 pg of GM-CSF per 150 mg of tissue, which approximately contained 106 of eosinophils, in the supernatant of NP explants for 24 h without any stimulation. Contents of both IL-5 and IL-3 were much less. We further studied survival of PB-EOS exposed to rhGM-CSF and found that 10(-6) M of BUD could only inhibit by less than one third the survival of PB-EOS exposed to an amount of rhGM-CSF, similar to that detected in the supernatant of NP explants. In addition, we exposed PB-EOS to 200 pg/ml of rhGM-CSF for a relatively long period of time (4 days) in order to mimic chronic exposure in the tissue and found that the survival of these cells was prolonged to the extent similar to that observed in NP-EOS. Our data suggests that the prolonged spontaneous survival of NP-EOS ex vivo is likely the result of sustained in vivo exposure to GM-CSF and budesonide has a smaller inhibitory effect in the survival of these eosinophils as compared to those from peripheral blood.

[Analysis of tympanoplasty at Chiba-Hokuso Hospital, Nippon Medical School].

We evaluated tympanoplasty statistically at Chiba-Hokuso Hospital, a branch hospital of Nippon Medical School. Especially, we discussed the improvement of auditory thresholds after operation, the type of tympanoplasty in valved, the availability of patch tests and complications.

Effects of prenatal methylazoxymethanol treatment on striatal dopaminergic systems in rat brain.

To further examine the effects of prenatal methylazoxymethanol (MAM) treatment on striatal dopaminergic systems, the status of presynaptic dopamine transporters was examined by quantitative autoradiography of [3H]GBR 12935 binding. Significantly higher [3H]GBR 12935 binding was seen in MAM-lesioned striatum in comparison to the controls, indicating relative dopaminergic hyperinnervation in MAM-induced hypoplastic striatum. The effect of prenatal MAM treatment on extracellular levels of dopamine and its metabolites in the striatum was also examined using in vivo microdialysis. As measured in conscious freely-moving rats, prenatal MAM treatment significantly increased basal dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) release in the striatum in comparison with control rats. These data suggest that in accordance with morphological dopaminergic hyperinnervation, dopaminergic functions are significantly augmented in MAM-lesioned brains. Thus, it is suggested that MAM-induced microencephalic rats should serve as a good animal model for the study of augmented dopaminergic functions in the striatum.

Effect of chronic amitriptyline administration on serotonergic receptors in rats with methylazoxymethanol-induced microencephaly.

Methylazoxymethanol (MAM)-induced cortical hypoplasia resulted in a 20% decrease in the Bmax of 5-HT2A receptors in the frontal cortex with no change in the Bmax of 5-HT1A receptors. Chronic treatment with amitriptyline did not further decrease the Bmax of 5-HT2A receptors in the MAM-lesioned cortex, suggesting that the persistent down-regulation of cortical 5-HT2A receptors in MAM-lesioned rats was induced by serotonergic hyperinnervation.

GM-CSF, IL-8, IL-1R, TNF-alpha R, and HLA-DR in nasal epithelial cells in allergic rhinitis.

Epithelial cells potentially contribute to airways inflammation by antigen presentation and the production of proinflammatory cytokines. This study investigated the immunocytochemical localization of interleukin-8 (IL-8), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 receptor (IL-1R Type I), tumor necrosis factor-alpha receptor (TNF-alpha R; 55kD), and human leukocyte antigen-DR (HLA-DR) on epithelial cells obtained by nasal brushing from 10 patients with allergic rhinitis in season and 15 healthy, nonallergic subjects. Six of the 15 nonallergic asymptomatic subjects had macroscopic evidence of nasal mucosal inflammation, and their brushings contained more than 10% neutrophils ("subclinical inflammation"). In normal control subjects, 8 +/- 7.5% of epithelial cells stained for HLA-DR, approximately one quarter stained for IL-8 and GM-CSF, and about one third stained positive for IL-1R and TNF-alpha R. The findings in subjects with allergic rhinitis in season and with subclinical neutrophilia were similar, and the numbers of cells staining for HLA-DR expression correlated with both neutrophil and lymphocyte content. These findings further support the conclusion that epithelial cells can contribute to inflammatory processes in the nasal mucosa. The findings emphasize the need to identify asymptomatic nasal mucosal inflammation in studies of the nasal mucosa.

Distinct immunohistochemical localization of IL-4 in human inflamed airway tissues. IL-4 is localized to eosinophils in vivo and is released by peripheral blood eosinophils.

Nasal polyposis and asthma are inflammatory conditions of the airways characterized by infiltration of activated inflammatory cells, particularly eosinophils. IL-4 is a multifunctional cytokine considered to play an important role in eosinophilic inflammation. We examined the cellular distribution of immunoreactive IL-4 in nasal polyps, as well as in the bronchial mucosa of both nonasthmatic control subjects (n = 6) and patients with well-characterized mild asthma (n = 6) subjected to a diluent or an allergen challenge. To determine eosinophilic contribution, tissue sections were counterstained with FITC after IL-4 immunostaining. No eosinophils were observed in the bronchial mucosa of nonasthmatic subjects. Nasal polyp tissues contained approximately 15 times more eosinophils per mm2 compared with bronchial tissues from asthmatics after a diluent challenge. Allergen challenge resulted in a marked increase in eosinophil density in bronchial tissues. A negligible number of cells immunostaining IL-4 was observed in bronchial tissues from nonasthmatic control subjects. The density of IL-4-positive cells in nasal polyp tissues was almost three times greater compared with asthmatics bronchial tissues after a diluent challenge. Approximately 90% of the IL-4-positive cells in bronchial tissues did not exhibit fluorescence after FITC counterstaining; in contrast, about 80% of the IL-4-positive cells in nasal polyp tissues did. We also show that peripheral blood eosinophils from allergic subjects express IL-4 mRNA by Northern blot analysis, particularly on stimulation with secretory IgA immune complexes. Finally, the supernatant of stimulated eosinophils contained approximately 50 pg/10(6) cells of IL-4 as determined by ELISA. These data demonstrate that eosinophils express the message and release IL-4 in vitro, and that these cells are the primary source of immunoreactive IL-4 in tissues undergoing chronic severe mucosal inflammation.

Triadic discrimination testing: refinement of Thurstonian and sequential sensitivity analysis approaches.

The effect of cognitive search strategies and variations in the oral environment on discrimination test performance were investigated. Subjects were required to discriminate between low concentration NaCl solutions and water using the 3-AFC and triangle test protocols. As predicted by Thurstonian modeling, subjects obtained a higher proportion of correct tests for the 3-AFC protocol than for the triangle protocol. The d' values obtained from both protocols corresponded. As predicted by the Sequential Sensitivity Analysis Model, which is largely based on changes in the oral environment, subjects obtained a higher proportion of correct tests for triads containing one NaCl stimulus than for triads containing one water stimulus. Measurement of physical signal strengths of the stimuli, by analysing the Na cation concentration change in the oral environment on tasting, indicated that the classical Thurstone-Ura two-distribution model was insufficient. The strong carry-over effects in the chemical senses require a model based on more than two distributions. It was also noted that subjects did not always use the search strategy required for a given test protocol. The artifactual effects of strategy change during an experiment are discussed.

Interleukin 10 inhibits lipopolysaccharide-induced survival and cytokine production by human peripheral blood eosinophils.

In this study we have investigated the effects of interleukin 10 (IL-10) on human peripheral blood eosinophils stimulated with granulocyte/macrophage colony stimulating factor (GM-CSF) and lipopolysaccharide (LPS). We show that LPS was able to enhance eosinophil survival in a dose-dependent manner, as well as release of the cytokines GM-CSF, tumor necrosis factor alpha, and IL-8. LPS-induced eosinophil survival was largely inhibited by an anti-GM-CSF neutralizing antibody and completely blocked by polymyxin B, suggesting GM-CSF involvement in the survival enhancing mechanism and LPS specificity, respectively. IL-10 significantly inhibited survival of, and cytokine production from, eosinophils induced by LPS, but did not inhibit the survival induced by GM-CSF. These observations suggest a novel activation mechanism of eosinophils and, also, that IL-10 may participate in the regulation of diseases characterized by eosinophil infiltration.

[The role of cervical inputs in maintaining equilibrium with regard to age].

The purpose of this experiment is to clarify the role of cervical inputs in maintaining body balance in humans, especially with regard to age. The difference in body sway between with and without neck vibratory stimulation, was analyzed in 83 normal subjects aged 6 to 73. The results showed that the difference between with and without the neck vibratory stimulation was the weakest in those aged from 15 to 44. The difference tended to be stronger in children under 14 years of age and in subjects over 60 years, in contrast to adolescents and nonelderly adults. The neck vibratory stimulation had more effect on forward-backward body sway than on right-left sway, with increasing age. In conclusion, the role of cervical inputs in maintaining equilibrium may change with age, in terms of the quantity and quality of its work capacity function.

Effects of chronic methamphetamine treatment on the binding parameters of [3H]SCH 23390, a selective D1-dopamine receptor ligand, in the rat brain.

Following the determination of the binding characteristics of [3H]SCH 23390 using slide-mounted tissue sections of molded minced forebrain, the effects of chronic methamphetamine (MAP) treatment (at a dose of 4 mg/kg for 14 days) on D1 receptors in the rat brain were studied using quantitative receptor autoradiography. There were no differences in both Kd(app) and Bmax values between vehicle- and MAP-treated rats, indicating that D1 receptors might not be involved in the development of behavioral sensitization in chronically MAP-treated rats.

Autosomal dominant type of endosteal hyperostosis with unusual manifestations of sclerosis of the jaw bones.

We report three cases of autosomal dominant type endosteal hyperostosis which occurred in one Japanese family. A new pattern of sclerotic changes in the jaw bones is evident. In all members of the family there was a symmetrical thickening of the diaphyseal cortices of the long bones. The affected bones were only minimally widened and the epiphyses and metaphyses were spared. Endosteal sclerosis of the neurocranium was present with loss of the diploë. The sclerotic changes included enlargement and mottled sclerosis of both the maxilla and mandible, with multiple embedded teeth and odontomas. The ramus of the mandible was spared. Severe sclerosis of the jaw bones was present only in a 28-year-old woman. The 2-year-son showed only focal sclerosis in the mandible, and his grandmother had minimal changes in the skeleton.

Time-dependent changes in the binding parameters of muscarinic cholinergic receptors in the rat brain.

Using quantitative receptor autoradiography, effects of several incubation times on the binding parameters for [3H]quinuclidinyl benzilate [(3H]QNB) binding were investigated in 5 discrete regions of the rat brain. There were no differences in Bmax-values between 3 incubation times. On the other hand, Kd(app)-values markedly depended on the duration of incubation time. Furthermore, Scatchard plots at low [3H]QNB concentrations showed an abnormal binding behavior, i.e. deviation from the straight line representing a single population of the binding site.

Quantitative autoradiography of muscarinic cholinergic receptors in the rat brain.

Using the in vitro autoradiographic technique with tritium-sensitive LKB sheet film and the liquid scintillation counting method, the distribution and the binding parameters of the muscarinic cholinergic receptors (MChR) were determined in various discrete regions of the rat brain. The results obtained in the present study were as follows: (1) Specific binding of [3H]QNB to the slide-mounted tissue sections increased slowly when incubated at room temperature; saturation occurred 2 h after incubation. Only 23% of [3H]QNB bound to the tissue section was dissociated 5 h after the addition of 20 microM atropine to the medium. These findings were very different from those obtained in the study using the tissue homogenates. (2) The regional distribution of MChR was determined using both autoradiographic and liquid scintillation counting methods. The distribution of MChR was heterogeneous, with highest densities in the striatum and nucleus accumbens and lowest in the globus pallidus, nucleus interpeduncularis and nucleus septi. Moreover, MChR were unevenly distributed within the subfields of each region. (3) In saturation binding studies using the slide-mounted tissue sections of 20 micron thickness the (Kd)app-values were similar but not exactly identical in 5 discrete regions, i.e. the striatum, somatosensory cortex, hippocampus (the subiculum + CA1 field), nucleus accumbens and gyrus dentatus, determined in the present study. The (Kd)app-value of each region was about 700 pM which was about 20 times higher than that obtained in the study using the tissue homogenates. However (Kd)app-values obtained with 5 and 10 micron tissue sections were approximately 3-fold lower.