Modulatory effects of serotonin on glutamatergic synaptic transmission and long-term depression in the deep cerebellar nuclei.

The deep cerebellar nuclei (DCN) are the terminal components of the cerebellar circuitry and constitute its primary output structure. Their activity is important for certain forms of motor learning as well as generation and control of movement. DCN neurons receive glutamatergic excitatory inputs from the pontine nuclei via mossy fibres (MFs) and concomitantly receive inputs from 5-HT-containing neurons of the raphe nuclei. We aimed to explore the roles of 5-HT at MF-DCN synapses by using cerebellar slices from 11 to 15-day-old rats. Bath application of 5-HT reversibly decreased the amplitude of stimulation-evoked excitatory postsynaptic currents (eEPSCs) via the activation of 5-HT1B receptors at the presynaptic terminals of the MFs. Burst stimulation of the MFs elicited long-term depression (LTD) at the MF-DCN synapses that require activation of the group I metabotropic glutamate receptor (mGluR). In the presence of 5-HT, the extent of burst-induced LTD of MF EPSCs was significantly reduced. Application of 5-HT also decreased the amplitude of mGluR-dependent slow EPSCs evoked by similar burst stimulation. Furthermore, (S)-3,5-dihydroxyphenylglycine (DHPG), a group I mGluR agonist, induced chemical LTD of MF EPSCs, and 5-HT had no significant effect on this LTD. Taken together, the results suggest that 5-HT not only has transitory inhibitory effects on MF EPSCs but also plays a role in regulating the long-term synaptic efficacy.

Prevention by lansoprazole, a proton pump inhibitor, of indomethacin -induced small intestinal ulceration in rats through induction of heme oxygenase-1.

The effect of lansoprazole, a proton pump inhibitor (PPI), on indomethacin-induced small intestinal ulceration was examined in rats, particularly in relation to heme oxygenase (HO)-1. The animals were administered indomethacin (10 mg/kg, p.o.) and killed 24 h later. Lansoprazole (30-100 mg/kg, p.o.) and omeprazole (30-100 mg/kg, p.o.) were given 30 min before the administration of indomethacin, while tin-protoporphyrin IX (SnPP: 30 mg/kg, i.v.), an inhibitor of HO-1, was injected 10 min before indomethacin or lansoprazole. Indomethacin produced hemorrhagic lesions in the small intestine, accompanied with an increase of mucosal invasion of enterobacteria, inducible nitric oxide synthase (iNOS) expression, and myeloperoxidase (MPO) activity in the mucosa. Pretreatment with lansoprazole dose- dependently reduced the severity of the indomethacin-induced intestinal lesions, with suppression of the increased MPO activity, while omeprazole had no effect. Pretreatment with SnPP significantly exacerbated these intestinal lesions and almost totally abolished the protective effect of lansoprazole. The up-regulation of iNOS mRNA expression following indomethacin was suppressed by lansoprazole in a SnPP-inhibitable manner, although the enhanced enterobacterial invasion remained unaffected. The amount of HO-1 protein in the intestinal mucosa was significantly increased by lansoprazole but not by omeprazole. Prior administration of carbon monoxide (CO)-releasing molecule-2 (CORM-2; 10 mg/kg, i.p.) significantly reduced the severity of these lesions and the enhancement of mucosal iNOS mRNA expression induced in the small intestine by indomethacin. These results suggest that lansoprazole prevents indomethacin-induced small intestinal ulceration, and this effect is associated with inhibition of iNOS expression, through up-regulation of HO-1/CO production in the mucosa.

Effects of nimesulide, a cyclooxygenase-2 selective inhibitor, on colitis induced tumors.

Cyclooxygenase-2 (COX-2) is known to suppress sporadic colorectal cancer, but effect of selective COX-2 inhibitor in UC-associated neoplasia is still unknown. This study investigated effect of a selective COX-2 inhibitor on colorectal carcinogenesis in experimental murine UC. Chronic colitis was induced in mice by four cycles of administration of dextran sulfate sodium (DSS) (i. e., 5 % DSS for 7 days and distilled water for the following 14 days), and the mice were sacrificed 120 days after the end of the fourth cycle. The mice were divided into the following five groups: Group A, served as a disease control; Group B, received a diet mixed with 400 ppm of nimesulide (NIM), a selective COX-2 inhibitor, during the whole period; Group C, received NIM during the four cycles of DSS administration; Group D, received NIM for 120 days from the end of the fourth cycle; Group E, served as a normal control. In Group D, NIM significantly suppressed the occurrence of dysplasia and/or cancer. The results show that NIM inhibited both dysplasia and cancer in DSS-treated mice, thus showing that NIM has preventive effects on the remission phase of carcinogenesis.

Endoscopic diagnosis of refractory ulcerative colitis.

BACKGROUND & AIM: To make the endoscopic characteristics and clinical appearance of refractory ulcerative colitis (UC) clear, we studied the frequency and location of refractory lesions of severe UC patients. METHODS: The subjects were a total of 68 patients with severe UC. 41 patients were identified with refractory UC, defined as poor response to high-dose systemic steroids (SH-resistant UC), and another group of 27 patients had non-refractory UC (SH-responsive UC). Two groups were compared by the following item; endoscopic findings, locations, effect of treatment, and relation to CMV infection. RESULTS: In SH-resistant UC, longitudinal ulcer and extensive mucosal abrasion were found with high frequency, and there were refractory lesion significantly in proximal colon. In case of UC with refractory lesion, there is a high possibility that treatment is ineffective. Of 14 UC patients with refractory lesion treated with LCAP, 10 obtained remission, whereas only 12 of 30 those patients with treated by only steroids achieved remission. Of 11 steroid-refractory UC patients with CMV detected, were refractory to steroids and had undergone. CONCLUSIONS: The findings of this study suggest that it is very important to decide on a patient's medication by relying on exact diagnosis concerning the condition of UC by endoscopy.

Expression of the EP4 prostaglandin E2 receptor subtype with rat dextran sodium sulphate colitis: colitis suppression by a selective agonist, ONO-AE1-329.

Expression of the EP4 receptor, a prostaglandin (PG)E2 receptor subtype, as well as disease suppression by the administration of a selective EP4 agonist (ONO-AE1-329) was investigated in the colorectal mucosa of rats with dextran sodium sulphate (DSS)-induced colitis. Rats were given drinking water containing 3% DSS for 2 weeks. Expression of EP4 receptor mRNA was barely detectable under normal conditions according to reverse transcription-polymerase chain reaction (RT-PCR). By 1 week after the initial administration of DSS, the receptor mRNA was strongly expressed. After ONO-AE1-329 was administered intracolonically to rats with DSS colitis for 7 consecutive days, erosion and ulceration decreased. Peripheral white blood cell (WBC) counts became less elevated. Interleukin (IL)-1beta and growth-regulated gene product/cytokine-induced neutrophil chemoattractant (GRO/CINC-1) concentrations in colorectal mucosa were lower than in colitis control group (IL-1beta: 12.8 +/- 4.6 and 30.8 +/- 6.2 microg/mg protein, P < 0.05; GRO/CINC-1: 15.5 +/- 3.0 and 39.2 +/- 5.4 microg/mg protein, P < 0.05), and the expression of the corresponding cytokine mRNA was strongly suppressed. IL-10 concentration was higher than in control group (14.5 +/- 1.7 and 7.9 +/- 1.2 microg/mg, P < 0.05), and the mRNA was more strongly expressed. These results suggest that the EP4 receptor is important in colonic inflammation, and that PGE2 suppresses DSS colitis at least partly via the EP4 receptor and the above cytokine changes. Intracolonic administration of selective EP4 agonist might have therapeutic applicability in inflammatory bowel disease such as ulcerative colitis.

Estimation of mucosal inflammatory mediators in rat DSS-induced colitis. Possible role of PGE(2) in protection against mucosal damage.

In order to investigate the mucosal injury mechanism in UC, we made dextran sulfate sodium (DSS)-induced colitis in rat and examined pathological findings, MPO activity, PGE(2) level, and local mRNA expression and secretion of IL-1 beta, TNF-alpha, GRO/CINC-1 and IL-10 in DSS colitis mucosa. Moreover, we estimated the correlation between the severity of mucosal damage and changes of these local inflammatory mediators' values. Neutrophil infiltration was marked and MPO activity was locally increased in proportion to the severity of mucosal damage. The mRNA expression and secretion of IL-1 beta, GRO/CINC-1 and IL-10 were increased. Especially, the secretions of IL-1 beta and GRO/CINC-1 were increased in proportion to the severity of mucosal damage. However, those of TNF-alpha were not increased in the colitis mucosa. An abnormal macrophage function and the presence of macrophage subtypes producing different cytokines would be predicted from our TNF-alpha data. The lesion was less severe in the colonic mucosa with higher levels of endogenous PGE(2), while it was more severe in the colonic mucosa with lower levels of endogenous PGE(2), implicating this compound as an inhibitory factor against the development of inflammation in the affected mucosa. Our results suggest that PGE(2) might have therapeutic applicability to UC.

Enprostil, a prostaglandin-E(2) analogue, inhibits interleukin-8 production of human colonic epithelial cell lines.

We previously reported that intracolonic administration of enprostil, a prostaglandin-E(2) (PGE(2)) analogue, had therapeutic effects on acute colitis induced in rodents by dextran sulfate sodium (DSS). In addition, production of growth-regulated gene product/cytokine-induced neutrophil chemoattractant-1 [GRO/CINC-1; an interleukin(IL)-8 like cytokine] was suppressed in the inflamed tissues. In the present study we used a human colon cancer cell line (HT-29) to investigate enprostil effects on the IL-8 production of intestinal epithelial cells stimulated by various stimulants. In a MTT assay, concentrations of enprostil >10(-5)M had cytotoxitic effects on HT-29 cells. Furthermore, 10(-6) M enprostil suppressed IL-8 production in HT-29 cells, SW620 and CaCo2 stimulated with interleukin-1 beta (IL-1 beta) or lipopolysaccharide (LPS), but did not suppress this response when cells were stimulated with tumour necrosis factor (TNF)-alpha. These results suggest that enprostil affects a point in the pathway between the IL-1 receptor or LPS receptor and nuclear factor-kappa B(NF-kappa B), without affecting the pathway between the TNF receptor and NF-kappa B, with the latter factor being required for the IL-8 gene transcription. The therapeutic effect of exogenous enprostil on DSS colitis may involve the inhibition of IL-8 production in colonic epithelial cells stimulated by IL-1 beta or LPS.

Therapeutic effect of intracolonically administered nuclear factor kappa B (p65) antisense oligonucleotide on mouse dextran sulphate sodium (DSS)-induced colitis.

Cytokines such as IL-1, tumour necrosis factor-alpha (TNF-alpha), IL-6 and IL-8 are increased in inflamed colonic mucosa after administration of mouse DSS. Nuclear factor kappaB (NF-kappaB) is a transcription factor which regulates the expression of these cytokine genes. The effect of intracolonically administered NF-kappaB (p65) antisense phosphorothioate oligonucleotide was examined in mouse DSS-induced colitis using drinking water containing 5% DSS. When antisense oligonucleotide was given on day 0, the disease activity index (DAI) representing clinical symptoms improved and the histological score decreased; furthermore, IL-1, IL-6, and TNF-alpha concentrations in rectal mucosa were lower compared with the control group. Clinical and histological improvement was also observed when antisense oligonucleotide was begun on day 2 but not on day 7. In addition, the distribution of antisense oligonucleotides was investigated by confocal laser microscopy. In colonic mucosa, oligonucleotides were predominantly localized to cells in the lamina propria, but also in the epithelium. Western blot analysis using homogenized rectal mucosa showed the decreased expression of NF-kappaB p65 in the antisense oligonucleotide-treated group, although it was increased in the colitis group. These results suggest that intracolonic administration of NF-kappaB antisense oligonucleotide may be effective in ulcerative colitis.

Prostaglandin E2 inhibits lesion formation in dextran sodium sulphate-induced colitis in rats and reduces the levels of mucosal inflammatory cytokines.

Effects of rectally injected prostaglandin E2 (PGE2) in rats with dextran sodium sulphate (DSS)-induced colitis were investigated in terms of histopathology, local myeloperoxidase (MPO) activity, local mRNA expression of interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and growth-regulated gene produced/cytokine-induced neutrophil chemoattractant (GRO/CINC)-1, and secretion of TNF-alpha and GRO/CINC-1. In animals with no PGE2 treatment, DSS-induced erosion and ulceration were particularly severe in the rectum and extended to the proximal colon. Neutrophil infiltration was characteristically present in the lesions and surrounding mucosa. MPO activity at lesion sites was increased. IL-1beta and GRO/CINC-1 mRNA expression was increased, while TNF-alpha mRNA expression was significantly decreased. GRO/CINC-1 secretion was increased but a similar elevation of TNF-alpha was not detected. In the PGE2-treated group, lesion formation was inhibited grossly and microscopically. Neutrophil infiltration and MPO activity in and around lesions were lessened. The reduction in TNF-alpha mRNA expression and secretion was not affected by PGE2. The expression of mRNA for IL-1beta and GRO/CINC-1 was reduced, as was the secretion of GRO/CINC-1. As mRNA expression and secretion of cytokines in lesions of non-PGE2-treated animals was similar to that reported in human ulcerative colitis, rectal injection of PGE2 may prove to be an effective therapy.

[Questionnaire on "having a family dentist" and the status of oral health].

In 1999, we gave a Questionnaire on "having a family dentist" to 414 workers (300 males, 114 females) at a government office. Oral health examination was also provided to the same subjects. The following results were obtained: 1. Female workers (43%) had a family dentist more often than male workers (29%), and the difference was significant (p < 0.01). 2. About 45% of the subjects received regular dental checkups at least once a year; however, half of them did not have a family dentist. 3. Convenience, comfort, and communication factors were the main reasons for choosing a particular dentist, while continuity, comprehensiveness, and specialty factors were not taken into consideration in choosing a dentist. 4. The persons who had family dentists had more FT and DMFT, and fewer total numbers of teeth than those without family dentists. From these results, it was suggested that people visited dentists with dental problems and received treatment repeatedly, and then they came to have family dentists. The habit of visiting a dentist regularly seeking checkups and prevention is not yet popular among Japanese adults. To improve oral health consciousness and to promote oral health, it should be recommended to receive regular checkups and oral health education by a family dentist. However, the present condition does not allow this system. Therefore in Japan, it should be encouraged to provide oral health examination and oral health education regularly at work sites, to improve the oral health of the adult population.

[Study on oral health status and health behavior of workers at government office].

The purpose of this study was to investigate the oral health status and health behavior of the adult population in the workplace. In 1998, oral health examination of 388 workers (male: 287, female: 101) at a government office was performed. A questionnaire was administered to obtain data regarding oral symptoms and health behavior. The results were as follows: Overall, 48% needed treatment for dental caries, 44% needed calculus removal, and 23% needed treatment for periodontal disease. Although there were no oral health complaints, 20% had early caries, 40% had dental calculus, and 19% had periodontitis. Compared to males, more females brushed their teeth, had home dentists (44%) and received more regular dental health check-ups at least once a year (48%). For males, those with home dentists had higher FT and DMFT in the twenties and thirties. There was no relationship between oral health status and regular check-ups in both males and females. The results revealed that receiving regular dental check-ups from home dentists was not popular in Japan. Further, the role of home dentists is not preventive oriented. It was concluded that it is necessary to provide regular oral health examination and health promotion programs for adult population at the workplace in Japan.

Inhibition of dextran sulphate sodium (DSS)-induced colitis in mice by intracolonically administered antibodies against adhesion molecules (endothelial leucocyte adhesion molecule-1 (ELAM-1) or intercellular adhesion molecule-1 (ICAM-1)).

We examined the effect of intracolonic administration of anti-adhesion molecule antibodies on DSS-induced colitis in mice. Immunohistochemical staining in mice with colitis showed increased expression of ELAM-1 and ICAM-1 on endothelial cells of vessels in the lamina propria and submucosa at sites of inflamed lesions. Intracolonic administration of anti-ELAM-1 or anti-ICAM-1 antibody decreased bloody stools, anaemia, and histologically evident damage, as well as myeloperoxidase activity and IL-1beta content. We concluded that adhesion molecule expression is important in the development of DSS-induced colitis in mice and that intracolonic administration of anti-adhesion molecule antibodies, especially anti-ELAM-1 antibody, effectively inhibits the colonic inflammation. Intracolonic administration of anti-adhesion molecule antibodies may show therapeutic promise in ulcerative colitis.

Positive correlation between levels of IL-1 or IL-2 and 1,25(OH)2D/25-OH-D ratio in synovial fluid of patients with rheumatoid arthritis.

The present study determined the levels in synovial fluid (SF) of vitamin D metabolites (1,25-dihydroxyvitamin D (1,25(OH)2D), 24,25-dihydroxyvitamin D (24,25(OH)2D) and 25-hydroxyvitamin D (25-OH-D)), and of the cytokines. We evaluated SF from 21 patients with rheumatoid arthritis (RA) and 6 patients with osteoarthritis (OA). The levels of vitamin D metabolites in SF, as determined by two different extraction methods, were significantly correlated (p < 0.05, n=7). The levels of 3 vitamin D metabolites were significantly higher in the RA SF than in OA SF (p < 0.05). The ratio of 1,25(OH)2D/25-OH-D in RA SF, which is presumed to reflect the activity of 25-OH-D-1-hydroxylase (1-OH-ase), was positively correlated with the levels of interleukin-1alpha (IL-1alpha), IL-1beta, and IL-2 in such SF, and was significantly higher than that in sera from RA patients. This suggests an important role for these cytokines in the activation of 1-OH-ase in RA synovium. The ratio of 24,25(OH)2D/25-OH-D, which is presumed to reflect 25-OH-D-24-hydroxylase (24-OH-ase) activity, was significantly correlated with 1,25(OH)2D levels only in RA SF, but not in sera from RA patients, suggesting a local regulation of vitamin D metabolism that 1,25-(OH)2D induces 24-OH-ase as in other target cells. Our observations suggested that 1,25(OH)2D and 24,25(OH)2D are produced locally from 25-OH-D in RA synovium, and that the syntheses of 1,25(OH)2D and 24,25(OH)2D may be affected by IL-1/IL-2 and 1,25(OH)2D in RA SF, respectively.

Case of a parathyroidectomized patient observed longitudinally by ultrasonography. Relationship between the growth rates and 1,25-dihydroxyvitamin D3 receptor contents in the parathyroid glands.

Parathyroid glands enlarge gradually with the progression of secondary hyperparathyroidism. The significance of down-regulation of the 1,25-dihydroxyvitamin D receptor (VDR) in parathyroid glands has been emphasized. Here we report a case in whom the relationship between the growth rates of the parathyroid glands and their VDR content was examined. A 36-year-old man, who had been hemodialyzed for 8.8 years because of chronic renal failure due to chronic glomerulonephritis, developed severe secondary hyperparathyroidism. The first ultrasonographic examination of the parathyroid glands, performed 10 months before parathyroidectomy, revealed that the sizes of the right upper (RU) and left upper (LU) glands were 10 x 8 x 5 and 14 x 10 x 9 mm3, respectively, although the right lower (RL) and left lower (LL) glands were not detected. The second ultrasonographic examination performed 5 days before PTX revealed that the RU gland had enlarged up to 24 x 12 x 10 mm3, while the LU gland remained unchanged at 16 x 9 x 8 mm3. At this time, the sizes of the RL and LL glands were determined only in the longitudinal section to be 10 x 5 and 4 x 3 mm2, respectively. In the excised specimens, the sizes and weights of the RU and RL glands were 25 x 10 x 9 mm3 and 1,950 mg and 17 x 10 x 8 mm3 and 1,160 mg, respectively, while those of the LU and LL glands were 16 x 10 x 7 mm3 and 850 mg and 9 x 8 x 7 mm3 and 350 mg, respectively. Histopathologic study demonstrated that the RU and RL glands exhibited nodular hyperplasia, while the LU and LL glands exhibited diffuse hyperplasia. Using a ligand binding assay, the VDR content of the rapidly growing RU and RL glands were significantly reduced to 32.6 +/- 9.6 and 32.7 +/- 5.2 fmol/mg protein, respectively, as compared to that of the LU gland with no significant proliferating activity (111.8 +/- 0.8 fmol/mg protein). It is of great interest that the smallest LL gland, which showed some proliferating potential in spite of a histologic pattern of diffuse hyperplasia, has a VDR content of 41.0 +/- 2.6 fmol/mg protein. In summary, it was implied from this case that the VDR content in the parathyroid gland might reduce as the growth rate of the parathyroid gland increases and, furthermore, that the VDR content seems to depend to some degree on the histopathologic pattern rather than on gland weight.

Determination of nadolol in serum by high-performance liquid chromatography with fluorimetric detection.

A sensitive high-performance liquid chromatographic method for a routine assay of nadolol in serum is described. Serum samples spiked with atenolol (internal standard) were extracted with diethyl ether. After centrifugation, the organic layer was evaporated to dryness. The residue was redissolved in the mobile phase and injected onto an octadecyl silica column (150 mm x 4.6 mm I.D.). The mobile phase was 0.05 M ammonium acetate (pH 4.5)-acetonitrile (85:15, v/v). Fluorometric detection (excitation 230 nm, emission 300 nm) was used. The minimum detectable level of nadolol in serum was 1 ng/ml.