Effects of vasopressin on histamine H(1) receptor antagonist-induced spatial memory deficits in rats.

The effects of [Arg(8)] vasopressin on histamine H(1) receptor antagonist-induced memory deficits were investigated using the eight-arm radial maze performance test in rats. Pyrilamine and diphenhydramine as well as scopolamine induced memory deficits characterized by increases in the number of total errors, reference memory errors and working memory errors. [Arg(8)] vasopressin improved not only scopolamine--but also pyrilamine--and diphenhydramine-induced memory deficits, although a high dose of [Arg(8)] vasopressin was needed to antagonize pyrilamine-induced memory deficits. The effects of pyrilamine on the brain [Arg(8)] vasopressin content were studied, and the hippocampus [Arg(8)] vasopressin content was shown to be decreased after pyrilamine injection. From these observations, it seems likely that [Arg(8)] vasopressin participates in not only the cholinergic system but also the histaminergic system in spatial memory.

Effects of fujibitol, a remedy for nasal symptoms, on experimental allergic rhinitis in rats.

The effects of Fujibitol, a preparation of crude drugs in wide clinical use for treatment of chronic rhinitis and empyema, on experimental allergic rhinitis in rats were studied. Fujibitol inhibited nasal allergic symptoms, i.e. sneezing and nasal rubbing, induced by antigen in sensitized animals. An increase in dye leakage into the nasal cavity induced by antigen was also inhibited by Fujibitol. On the other hand, no inhibitory effects were observed on either the nasal allergic symptoms or increase in dye leakage into the nasal cavity induced by histamine. However, Fujibitol was effective in inhibiting histamine release from the nasal cavity induced by antigen. Oxatomide used as positive control drug showed potent inhibitory effects on nasal symptoms and dye leakage into the nasal cavity induced by histamine and antigen. These results suggested that Fujibitol showed a remarkable protective effect against experimental rhinitis induced by antigen via inhibition of histamine release from the nasal cavity.

Low novelty-seeking differentiates obsessive-compulsive disorder from major depression.

OBJECTIVE: To make a direct comparison of patients with obsessive-compulsive disorder (OCD) and major depression (MD) and a normal control group in terms of the Temperament and Character Inventory (TCI) personality dimensions. METHOD: Additionally to 43 patients with primary OCD, 43 MD patients and 43 normal subjects who were matched against the OCD patients for sex and age filled out the TCI. RESULTS: Compared to the controls, the OCD and MD patients scored significantly higher on harm avoidance and significantly lower on self-directedness and co-operativeness. The OCD patients scored significantly lower on novelty-seeking than the MD patients and the controls. CONCLUSION: Whereas OCD and MD share similar personality deviations on harm avoidance, self-directedness and co-operativeness, OCD is distinguishable from MD in terms of low novelty-seeking. Low novelty-seeking may have a profound relationship to the specific aetiology of OCD.

Clinical investigation of the ICD-10 subcategories for obsessive-compulsive disorder.

To examine the validity of ICD-10 subcategories for obsessive-compulsive disorder (OCD), the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) was applied to 53 OCD patients. The ratios of Y-BOCS compulsions subscore to obsessions subscore were calculated. The group with ratios around one consisted of patients diagnosed in three subcategories (F42.0, F42.1 and F42.2). This suggests that subjective subcategorization such as ICD-10 may be inadequate to differentiate between predominantly obsessive and compulsive patients compared with differentiation by quantitative assessment such as the Y-BOCS ratio. Thus, in selecting the appropriate therapeutic methods, we emphasize the usefulness of quantitative assessment in clinical settings.

Stac, a novel neuron-specific protein with cysteine-rich and SH3 domains.

In RLGS-M (restriction landmark genomic scanning using methylation-sensitive endonuclease) displays of mouse brains, spot #91 is one of tissue-specific gel spots whose intensity changes developmentally. We have now cloned the corresponding cDNA from this spot and analyzed its structure and expression. The deduced amino acid sequence revealed that the #91 cDNA encodes a novel protein of 403 amino acids which consists of a cysteine-rich domain and a SH3 domain. We designated this gene as Stac. Northern blotting and in situ hybridization analyses demonstrated that 2.7 kb of Stac mRNA is expressed predominantly in brain and neurons, especially in hippocampus, cerebellum and inferior olive. Further, the gene product of 47 kDa was found by western blotting analysis in the soluble fractions of brain as well as Stac-expression vector-transfected NIH3T3 cells. Although the function of Stac is unknown so far, it is likely involved in a neuron-specific signal transduction.

Correspondence of RLGS-M spot behavior with tissue expression on mouse homologue of DP1/TB2 gene.

RLGS-M is a novel method which enables visual and quantitative detection of many methylatable loci as two-dimensional gel spots. We isolated and characterized a mouse genomic clone corresponding to the RLGS-M gel spot #109v, which showed different intensities between two tissues: the spot was much more faint in liver than in brain. Southern and Northern blot analysis disclosed this spot behavior to be related to the methylation state of NotI site and also to be correlated with the differential expression in both tissues; the transcripts for #109v were observed in the brain but hardly any in the liver. Furthermore, in the base sequence analysis, this #109v gene was found to be the mouse homologue of human DP1/TB2 gene and had a CpG island adjacent to the NotI site. These results indicate that the mouse DP1/TB2 gene is expressed corresponding to its RLGS-M spot behavior which is related to its methylation condition.

Reliability and validity of the Japanese version of the Yale-Brown Obsessive-Compulsive Scale.

The reliability and validity of the Japanese version of the Yale-Brown Obsessive-Compulsive Scale (JY-BOCS) were determined by 20 raters for 12 Japanese patients with obsessive compulsive disorder at four institutions. Interrater reliability for the total JY-BOCS score was excellent, and the intraclass correlation coefficient was high (ICC = 0.960). Internal consistency was also excellent (Cronbach's alpha = 0.889). Concurrent and discriminant validity of the JY-BOCS was examined by comparing the scores on the JY-BOCS with those on the Maudsley Obsessional Compulsive Inventory (MOCI) and scales for depression and anxiety. A slight correlation was found between scores on the JY-BOCS and MOCI, but no significant correlations were found between scores on the JY-BOCS and those on scales for depression or anxiety.

A spot cloning method for restriction landmark genomic scanning.

We introduce two new methods for target cloning of DNA fragments corresponding to spots on the two-dimensional profile of restriction landmark genomic scanning (RLGS). One is a restriction trapper-based method and the other is a polymerase chain reaction (PCR) mediated method. Both are designed to select the target DNA fragments from a large amount of unlabeled background DNA fragments in the RLGS gel which produce background clones. The restriction trapper method is simple, with a cloning efficiency that is not biased by the length of the target DNA nor by its GC content. On the other hand, the PCR-mediated method is efficient for cloning DNA fragments from a small amount of starting materials. These methods provide us with powerful tools for isolating DNA clones identified by the RLGS system as interesting spots. This paper reports the precise protocols of these methods and discusses their application and usefulness.

Accessibility to tissue-specific genes from methylation profiles of mouse brain genomic DNA.

The DNA methylation status of a large number of genomic loci is visualized simultaneously and quantitatively as two-dimensional gel spots in the newly developed restriction landmark genomic scanning with a methylation-sensitive restriction enzyme (RLGS-M). Here, we demonstrate that RLGS-M using NorI as a methylation-sensitive enzyme could also scan gene loci of mammalian genomes, since almost all of the NotI loci corresponding to randomly chosen RLGS-M spots were located near or in transcriptional units (6 out of 7 NotI-linking clones) when mouse brain genomic DNA was used. This supports the previous prediction that most NotI sites are located in CpG islands (Lindsay and Bird, Nature 1987, 327, 336-338). Furthermore, beginning with RLGS-M spots we examined how to approach their corresponding RNA messages, whose expression may be associated with methylation. We compared RLGS-M patterns among various developmental stages of the mouse brain from embryonic day 9.5 to postnatal 8 weeks or among in vitro cell lines, and detected alterations of RLGS-M spots which were due to methylation of NotI sites. Two experiments using NotI-linking clones or polymerase chain reaction (PCR) were carried out to approach to their corresponding RNA messages. Consequently, we isolated two PCR-amplified clones (# 15 and # 91) which corresponded to methylatable loci and gave positive signals to mRNA from the adult brain. Furthermore, we identified two NotI-linking clones (C211 and C198) whose corresponding NotI loci localized near or at transcriptional units and were methylated in cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Genomic analysis of a NF1-related pseudogene on human chromosome 21.

A neurofibromatosis type-1 (NF1)-related locus on human chromosome 21 has been characterized. A detailed genomic mapping performed by yeast artificial chromosome (YAC) dot-hybridization revealed that the NF1-related locus is close to the sequence-tagged site (STS), D21S329 (G52E12), which is located on the proximal region of 21q11.2. Sequence analysis showed that this locus seemed to be conserved to the NF1 gene only in several partial regions. Two exon-like segments corresponding to exons 8 and 9 of NF1 were found, in addition to two previously found fragments corresponding to exons 7 and 11. Other exon-like segments were not found in the region so far sequenced. Comparing these homologous segments with the NF1 cDNA, a 2-bp deletion appeared in exon 8 of the locus, resulting in the existence of stop codons in all reading frames. In addition, there were no amplified fragments derived from the related locus by reverse transcription-PCR. Thus, our results suggest that the NF1-related locus on chromosome 21 is a nonprocessed pseudogene.

A PCR-mediated method for cloning spot DNA on restriction landmark genomic scanning (RLGS) gel.

A PCR-mediated direct cloning for target spot DNA from RLGS gel has been established. The method consists of PCR amplification of adaptor-ligated spot DNA fragments without excluding similar-sized DNA fragments co-localized on RLGS gel, and following selective ligation with the NotI-dT vector. Applying this method, we have successfully cloned several DNA fragments derived from target spots whose intensities change developmentally due to DNA methylation in the telencephalon of C3H/HeN mice. Since only a few micrograms of total DNA is sufficient for our spot cloning, our method may be highly useful when the total DNA sample prepared for cloning is limited.

An exon-trapping system with a newly constructed trapping vector pEXT2; its application to the proximal region of the human chromosome 21 long arm.

We have developed an exon-trapping system with a newly constructed trapping vector containing multiple cloning sites (designated pEXT2). The system revealed high sensitivity for trapping a control exon from several hundred kbp of DNA. We have applied the system to the cosmid clones located on human chromosome 21p11-q21, and identified two fragments highly homologous to neurofibromatosis 1 (NF1) gene and a clearly transcribed fragment hybridized with approximately 1.6 kb RNA from human brain and human glioblastoma A172 cell.

Effects of L-threo- and erythro-3,4-dihydroxyphenylserine on learning performance and concentrations of brain noradrenaline and its metabolites in rats.

Effects of L-threo and L-erythro-3,4-dihydroxyphenylserine [DOPS, precursor amino acids for noradrenaline (NA)] on the learning performance in a maze paradigm designed to model on the water maze paradigm using a multicomputerized behavioral analysis system were studied. A marked facilitation of learning performance was observed in rats after an intraventricular injection of 5 micrograms L-threo-DOPS (the s-NA precursor), and this effect was inhibited by a simultaneous administration of 1 or 2 micrograms propranolol (a beta-adrenergic antagonist). As concentrations of brain NA, 3-methoxy-4-hydroxyphenylglycol, and normethanephrine were increased by the injection of 5 micrograms L-threo-DOPS, the effect seemed to be derived from activation of beta-adrenoceptors in the CNS by the formed s-NA. On the other hand, an intraventricular injection of 5 micrograms L-erythro-DOPS (the r-NA precursor) attenuated the learning performance, and this effect was probably caused by the formed r-NA from L-erythro-DOPS.

Concentrations of beta-phenylethylamine in plasma and plateletes of schizophrenics.

The plasma and platelet PEA levels of 20 normal subjects and 17 schizophrenic patients were investigated using a high-performance liquid chromatography. In the normals the mean plasma and platelet levels of PEA were 4.9 +/- 1.9 ng/ml and 1.78 +/- 1.01 ng/mg protein, respectively, while in the schizophrenics, those were 12.1 +/- 7.9 ng/ml and 0.77 +/- 0.5 ng/mg protein, respectively. The plasma PEA levels of the schizophrenics were significantly higher than those of the normals, and the platelet PEA levels of the schizophrenics were lower than those of the normals.

Rapid and sensitive determination of beta-phenylethylamine in animal brains by high performance liquid chromatography with fluorometric detection.

A reversed phase HPLC method with fluorometric detection for the analysis of beta-phenylethylamine has been developed using p-methoxyphenylethylamine as an internal standard. Two columns, containing 200 microL of Dowex 50-X8 and Amberlite CG-50 respectively, were used to prepare a fraction containing beta-phenylethylamine. The recoveries of beta-phenylethylamine and p-methoxyphenylethylamine were 53.9 +/- 9.4% and 68.1 +/- 12.4%, respectively, and elution profile of p-methoxyphenylethylamine was sufficiently well correlated with that of beta-phenylethylamine. Regional distributions of beta-phenylethylamine in rat and mouse brains were determined. The highest concentrations were found in hypothalamus and hippocampus in both animals.

Studies on 3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxyphenylglycol (DHPG) levels in human urine, plasma and cerebrospinal fluids, and their significance in studies of depression.

Both concentrations of total 3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxyphenylglycol (DHPG) in the human urine, plasma and CSF were determined with a high-pressure liquid chromatography with electrochemical detection in order to clarify the dynamic change in these noradrenaline metabolites. Three different biological fluids were collected simultaneously from 16 orthopedic patients who were regarded clinically as substitutes for normal subjects. In the urine, the MHPG concentrations were 1.67 +/- 0.65 micrograms/mg creatinine (mean +/- S.D.) and DHPG 0.39 microgram/mg creatinine +/- 0.21. The plasma levels were 21.16 ng/ml +/- 9.58 for MHPG, and 19.58 ng/ml +/- 8.13 for DHPG. The CSF levels of MHPG and DHPG were 24.08 ng/ml +/- 8.10 and 34.76 ng/ml +/- 11.46, respectively. The CSF levels of these metabolites were correlated significantly with those in the plasma (r = 0.852, p less than 0.001 for MHPG; r = 0.799, p less than 0.001 for DHPG), while no significant correlations were found between the urinary levels and either the plasma or CSF levels of these metabolites. In the urine, the MHPG levels were proportional to the DHPG levels, while the former were inversely proportional to the latter in the plasma or CSF. Neither the MHPG nor DHPG levels in the urine from depressed patients revealed to have any significant correlation with their clinical assessments using the Hamilton Rating Scale Score (HRS). The patients were treated with an antidepressant active selectively on the noradrenergic system, and no significant changes in urinary excretion of these metabolites were observed before and after the drug treatment. These findings suggest that in the case of psychiatric disorders such as depression, these compound levels in the plasma or CSF would provide more important information than those in the urine.

Determination of beta-phenylethylamine concentrations in human plasma, platelets, and urine and in animal tissues by high-performance liquid chromatography with fluorometric detection.

A highly sensitive method for the determination of beta-phenylethylamine in human plasma, platelets, and urine and in mouse tissue is described. The method is based on a two-step isolation using cation-exchange columns followed by reverse phase high-performance liquid chromatography with fluorometric detection. The recovery of the amine through the whole procedure was almost complete, ranging from 99 to 101%. The calibration graph appeared linear over the range of 50 to 5000 pg/injection. Urinary excretion of beta-phenylethylamine in humans ranged from 0.93 to 51.20 ng/mg creatinine. The amine was also detectable in plasma and platelets. Of the various mouse tissues examined, the highest concentrations were found in the small intestine, followed by the blood and liver. Concentrations of about 5 ng/g wet wt were detected in brain tissue, which increased remarkably after inhibition of monoamine oxidase by pargyline.

[Studies on the reverse tolerance phenomenon after repeated methamphetamine treatment in rats].

Male rats of the Wistar strain were divided into three groups, to which three different doses of methamphetamine (MAP), 0.5 mg, 2 mg, and 4 mg/kg/day, respectively, were intraperitoneally administered 5 times per week for 36 days. Stereotyped behaviors were scored according to the method of Ellinwood & Balster. During the period of repeated MAP treatment, these behaviors increased in a dose dependent manner. Group I treated with 0.5 mg/kg of MAP demonstrated more prominently an increased sensitivity to MAP than the other two, Group II with 2 mg/kg and Group III with 4 mg/kg. Following the withdrawal period of 4 weeks, the rats were reinjected with 0.5 mg/kg of MAP. They showed higher scores of stereotypies than those treated acutely with MAP, that is, reverse tolerance for MAP was observed, although this phenomenon was not necessarily observed in a dose dependent manner for each variable of the behaviors. Lethal autonomic response was observed in Group I and II, in which animals appeared to have reverse tolerance for autonomic response to MAP reinjection in contrast to in Group III. It can be concluded that repeated administration of MAP induces stereotyped behaviors in a dose dependent manner, while autonomic response is not likely to form tolerance.