The roles of histamine, leukotriene C4 and bradykinin on nasal vascular permeability in experimental nasal allergy of guinea pigs.

The releases of histamine, leukotriene C4 (LTC4) and bradykinin into the nasal cavity were measured following nasal antigenic challenge in ovalbumin (OA) sensitized guinea pigs, or following nasal stimulation with one of these chemical mediators in OA-non-sensitized animals. In sensitized animals, increased vascular permeability of nasal mucosa was recognized immediately after antigenic stimulation and lasted for 90 minutes. The release of histamine into the nasal lavage fluid was observed only immediately after the antigenic stimulation. The releases of LTC4 and kinins into the nasal lavage fluid were augmented not only immediately after the antigenic challenge, but also 60 to 90 minutes after the stimulation. Nasal stimulation with one of these chemical mediators also increased nasal vascular permeability, but lasted for less than 40 minutes. These results suggest that the antigen-induced release of these chemical mediators might play some important roles in early increase of nasal vascular permeability, and that the increase of LTC4 and kinin levels might be involved in the prolonged nasal vascular permeability after nasal allergic response.     

Alterations of the nucleolar organizer regions during 4-nitroquinoline 1-oxide-induced tongue carcinogenesis in rats.

The number of silver-stained nucleolar proteins (AgNORs) was enumerated in preneoplastic and neoplastic rat tongue lesions induced by 4-nitroquinoline 1-oxide (4-NQO). Male ACI/N rats were given 0 to 10 p.p.m. 4-NQO for 12, 20 or 36 weeks to induce hyperplasia, dysplasia and neoplasm in tongue. The mean numbers of AgNORs stained by a modified one-step silver colloid method in various epithelial lesions were as follows: normal squamous epithelium (n = 5), 1.52 +/- 0.03; non-lesional squamous epithelium (n = 5), 1.58 +/- 0.04; hyperplasia (n = 20), 1.84 +/- 0.15; dysplasia (n = 20), 2.32 +/- 0.12; papilloma (n = 6), 2.23 +/- 0.10; and squamous cell carcinoma (n = 4), 3.06 +/- 0.26. Thus, the mean number of AgNORs showed a stepwise increase from untreated and treated, histologically normal squamous epithelium through hyperplasia and dysplasia to squamous cell papilloma and carcinoma, although the value of severe dysplasia was between those of papilloma and carcinoma. These results indicate that the mean number of AgNORs may reflect the proliferative nature of tongue lesions, as suggested in carcinogenesis of other organs, and also suggest that severe dysplasia is a direct precursor lesion for squamous cell carcinoma of the tongue induced by 4-NQO.     

Factors in the prognosis of early gastric cancer

Prognosis factors for 1,012 subjects who underwent gastrectomy for early gastric cancer were studied over a 21-year period from 1965 to 1985. We divided submucosal cancer (sm) into two subgroups, classifying the one as sm II-1 with small cancerous nests in the submucosa and the other as sm II-2. Our cases of early gastric cancer fell into the following groups: m(mucosal cancer), 451 cases; sm II-1, 106; sm II-2, 396 and multiple cancer, 59. The rate of lymph node metastasis was 1.1% in the m group, 5.7% in the sm II-1 group, 24.5% in the sm II-2 group and 6.8% in the multiple group. Gastrectomy with removal of the second-group lymph node proved adequate for sm II-2 group. The five year survival rate was 94.5% in the m group, 94.9% in sm II-1 group, 93.6% in the sm II-2 group and 91.7% in the multiple cancer group. The rate of recurrence in m group was 0.9%, against 3.3% for sm II-2 group. The sm II-2 with histologically well-differentiated type, n(+) and macroscopically Borrmann type are high-risk groups. Adjuvant chemotherapy is necessary to obtain improved surgical results for the high risk group. There was a total of 21 non-curative resection cases among which distant metastases were observed. Cases of early gastric cancer exist which are beyond help by surgical means.     

Aspirin test for differentiation of unilateral renovascular hypertension from hyperreninemic essential hypertension

Responses of renin release and blood pressure to aspirin DL-lysine (ASP) were examined to find out if the responses could help in the differentiation between unilateral renovascular hypertension (RVH) and hyperreninemic essential hypertension (EHT). The two studies involved ten patients with unilateral RVH, eight with hyperreninemic EHT, and five with hyporeninemic EHT. In a radiological study, before and 30 min after an intravenous injection of ASP (18 mg/kg), renal venous and abdominal aortic plasma was sampled and assayed for prostaglandin (PG) E2 and plasma renin activity (PRA). Systemic blood pressure was measured serially. The reproducibility of the responses to ASP was confirmed in a bedside study. In unilateral RVH, ASP suppressed renin release from the stenotic kidney and reduced the renal vein PRA ratio to less than 1.5 via the inhibition of PG synthesis, which is accelerated in that kidney. The mean suppression of aortic PRA at this dose of ASP was 35% in these patients, and their blood pressure decreased in proportion to the suppression of PRA. However, in the two EHT groups, ASP elevated the mean blood pressure. The renal synthesis of PGE2 was inhibited by ASP in all patients, but the suppression of PRA, while small, was significant (19% in the aorta) in the patients with hyperreninemic EHT, and not significant in patients with hyporeninemic EHT. The different responses of blood pressure and PRA to ASP between RVH and EHT were reproducible in the bedside study.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ghrelin directly regulates bone formation.

To clarify the role of ghrelin in bone metabolism, we examined the effect of ghrelin in vitro and in vivo. Ghrelin and its receptor, GHS-R1a, were identified in osteoblasts, and ghrelin promoted both proliferation and differentiation. Furthermore, ghrelin increased BMD in rats. Our results show that ghrelin directly affects bone formation. INTRODUCTION: Ghrelin is a gut peptide involved in growth hormone (GH) secretion and energy homeostasis. Recently, it has been reported that the adipocyte-derived hormone leptin, which also regulates energy homeostasis and opposes ghrelin's actions in energy homeostasis, plays a significant role in bone metabolism. This evidence implies that ghrelin may modulate bone metabolism; however, it has not been clarified. To study the role of ghrelin in skeletal integrity, we examined its effects on bone metabolism both in vitro and in vivo. MATERIALS AND METHODS: We measured the expression of ghrelin and growth hormone secretagogue receptor 1a (GHS-R1a) in rat osteoblasts using RT-PCR and immunohistochemistry (IHC). The effect of ghrelin on primary osteoblast-like cell proliferation was examined by recording changes in cell number and the level of DNA synthesis. Osteoblast differentiation markers (Runx2, collagen alpha1 type I [COLI], alkaline phosphatase [ALP], osteocalcin [OCN]) were analyzed using quantitative RT-PCR. We also examined calcium accumulation and ALP activity in osteoblast-like cells induced by ghrelin. Finally, to address the in vivo effects of ghrelin on bone metabolism, we examined the BMD of Sprague-Dawley (SD) rats and genetically GH-deficient, spontaneous dwarf rats (SDR). RESULTS: Ghrelin and GHS-R1a were identified in osteoblast-like cells. Ghrelin significantly increased osteoblast-like cell numbers and DNA synthesis in a dose-dependent manner. The proliferative effects of ghrelin were suppressed by [D-Lys(3)]-GHRP-6, an antagonist of GHS-R1a, in a dose-dependent manner. Furthermore, ghrelin increased the expression of osteoblast differentiation markers, ALP activity, and calcium accumulation in the matrix. Finally, ghrelin definitely increased BMD of both SD rats and SDRs. CONCLUSIONS: These observations show that ghrelin directly stimulates bone formation.     

Combined effects of cabergoline and L-dopa on parkinsonism in MPTP-treated cynomolgus monkeys

Summary The behavioral effects of L-dopa or cabergoline alone were compared with those of the joint administration of the two drugs in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned parkinsonian cynomolgus monkeys with attention to the induction of hyperactivity and dyskinesia. Cabergoline alone at 0.2mg/kg or less improved in a dose-dependent fashion the parkinsonism without inducing hyperactivity and dyskinesia following a single subcutaneous injection. L-dopa alone improved the parkinsonism, but induced hyperactivity and dyskinesia, depending on the dose applied. Doses required for 50% amelioration by L-dopa and cabergoline were 10 and 0.038mg/kg, s.c., respectively. With low doses (50%-amelioration doses), cabergoline or L-dopa alone improved the parkinsonism without induction of hyperactivity and dyskinesia, but the duration of action was brief. Cabergoline in combination with L-dopa was highly effective in improving motor disability without induction of hyperactivity and dyskinesia. Moreover, the duration of action was more prolonged with the coadministration than with the single administration of each drug. These findings suggest that the combined therapy with low doses of L-dopa and cabergoline is beneficial for treating patients with advanced Parkinson's disease.     

Preoperative combined treatment with radiation,intraluminal hyperthermia,and 5-fluorouracil suppositories for patients with rectal cancer

A clinical trial of a novel modality consisting of preoperative irradiation and intraluminal hyperthermia therapy, with or without 5-fluorouracil (5-FU) suppositories, was conducted on 30 patients with rectal cancer. To create hyperthermia in rectal cancer, an intraluminal electrode connected to a radiofrequency system was devised. Patients were treated preoperatively with a total irradiation dose of 30 Gy, being 3 Gy every 2 days, and 5 applications of hyperthermia given twice a week, with or without 5-FU suppositories, given as 100 mg a day to a total of 2,000 mg. Twenty patients received irradiation, intraluminal hyperthermia, and 5-FU suppositories, being the three-combination treatment, and the remaining 10 received irradiation and intraluminal hyperthermia, being the two-combination treatment. Shrinkage of the rectal cancer was observed macroscopically in 26 patients, resulting in a flattened cancerous crater or shallow ulcer. Microscopic examination revealed a marked decrease in the number of cancer cells. In fact, three resected specimens showed no evidence of cancer cells microscopically. These striking reductions in cancer size enabled us to successfully perform super-low anterior resections in four patients, with anastomoses being made just above the dentate line with a sufficiently safe margin. No serious complications were encountered in this series.     

Further simplification of ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-htlv-i igg using microplates and fluororeader

In a previously reported ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-HTLV-I IgG, polystyrene beads were handled with tweezers, and bound b?-D-galactosidase activity was measured with a fluorometer. The use of tweezers was causative of false-positivity by carryover, and testing many samples was difficult. Recently, these drawbacks have been minimized using microplates and a fluororeader. However, tweezers were still required in the initial and final steps. In the present study, the immune complex, comprising 2,4-dinitrophenyl-antigen, anti-HTLV-I IgG, and antigen-b?-D-galactosidase conjugate, was formed in and trapped onto microplate wells coated with (anti-2,4-dinitrophenyl group) IgG. Subsequently, the microplate wells were incubated with ?N-2,4-dinitrophenyl-L-lysine and polystyrene beads, modified by attaching to plates through cylindrical bars and coated with (antihuman IgG γgM-chain) IgG, to transfer the immune complex from the microplate wells to the modified polystyrene beads. Alternatively, modified polystyrene beads coated with (anti-2,4-dinitrophenyl group) IgG and microplate wells coated with (antihuman IgG γgM-chain) IgG were substituted for microplate wells coated with (anti-2,4-dinitrophenyl group) IgG and modified polystyrene beads coated with (antihuman IgG γgM-chain) IgG, respectively. The fluorescence intensity for bound b?-D-galactosidase activity was quickly measured with a fluororeader. Thus the modified polystyrene beads were transferred from wells to wells more quickly and easily without tweezers, eliminating false-positivity due to carryover, and it became easy to test many samples with high sensitivity and reliability, although the assay of bound b?-D-galactosidase activity became slightly more time-consuming.©1995 wiley-Liss, inc.     

Investigation into the 5-hydroxytryptamine-induced atropine-resistant neurogenic contraction of guinea-pig proximal colon.

1. The aim of this study was to characterize the receptors mediating the atropine-resistant neurogenic contraction to 5-hydroxytryptamine (5-HT) in the longitudinal muscle of the guinea-pig proximal colon and to determine the type of tachykinin receptors involved in the contractile response to 5-HT by the use of selective antagonists. 2. In the presence of atropine (0.3 microM), guanethidine (5 microM), hexamethonium (100 microM), ketanserin (0.1 microM) and indomethacin (3 microM), 5-HT (0.01-3 microM) produced concentration-dependent neurogenic contractions of colonic strips and at 0.3 microM produced a maximal effect (pEC50 = 7.39 +/- 0.09, n = 18). The 5-HT4 receptor stimulant, 5-methoxytryptamine (5-MeOT, 0.03-10 microM) also produced neurogenic contractions with similar maximum effect to those of 5-HT (pEC50 = 6.89 +/- 0.16). 3. The 5-HT4 receptor antagonist, DAU 6285 (3 microM) shifted the concentration-response curves to both 5-HT and 5-MeOT to the right without significant depression of the maximum, but the 5-HT1/5-HT2 receptor antagonist, metitepine (0.1 microM) and the 5-HT3 receptor antagonist, ondansetron (0.3 microM) had no effect on the control curves to 5-HT and 5-MeOT. 4. The selective NK1 receptor antagonist, FK 888 (1 microM) markedly attenuated the contractions to 5-HT and 5-MeOT. In contrast, the selective NK2 receptor antagonist, SR 48968 (10 nM) and the selective NK3 receptor antagonist, SR 142801 (10 nM) had no effect on the contractions to 5-HT and 5-MeOT. 5. These results indicate that the 5-HT-induced atropine-resistant neurogenic contraction of guinea-pig proximal colon is due to activation of 5-HT4 receptors, presumably located on excitatory motor neurones, innervating the longitudinal muscle. The contraction evoked by activation of the 5-HT4 receptors is mediated primarily via NK1 receptors but not NK2 or NK3, suggesting that the 5-HT4 receptor-mediated contraction is evoked indirectly via tachykinin release from tachykinin-releasing excitatory neurones.