Mapping of the critical region of mitogene-inducible gene-6 for NF-kappaB activation

Mitogene-inducible gene-6 (Mig-6) is involved in the intracellular signaling pathway as an adaptor molecule. Mig-6 expression is rapidly induced upon various kinds of mitogenic and stressful stimulations. We previously demonstrated that Mig-6 expression is induced by activated Ki-ras in human colon cancer cells and the limited proteolytic processed NH(2)-terminal region containing the Cdc42/Rac interaction and binding (CRIB) domain of Mig-6 is bound to IkappaBalpha, resulting in NF-kappaB activation. In this study, we tried to determine the critical region of Mig-6 for regulating NF-kappaB activation by using various kinds of deletion-constructs of Mig-6. The CRIB domain-deleted Mig-6 fragment (residues 68 to 462) did not show significant NF-kappaB activation. Furthermore, NH(2)-terminal 148, 158 and 180 amino acid regions of Mig-6 did not show NF-kappaB activation. On the other hand, the NH(2)-terminal 264 amino acid region of Mig-6 did show NF-kappaB activation to a similar extent of the full length of Mig-6. Interestingly, the full length and NH(2)-terminal 264 amino acid region of Mig-6 produced two kinds of cleaved NH(2)-terminal fragments, N1 and N2 with an approximate molecular size of 5 and 7 kDa, respectively. The NH(2)-terminal 148 and 158 amino acid regions of Mig-6 did not produce N1 or N2 fragments, while the NH(2)-terminal 180 amino acid region of Mig-6 only produced the N1 fragment. Furthermore, the N2 fragment was bound with IkappaBalpha, but the N1 fragment was not. These results together suggested that the NH(2)-terminal 264 amino acid region is critical for NF-kappaB activation and proper limited proteolytic processing of Mig-6.     

A case of 10-year survival after combination chemotherapy of 5'-deoxy-5-fluorouridine and cisplatin and adjuvant surgery for gastric cancer with liver metastasis

A 36-year-old woman was diagnosed with type 3 advanced gastric cancer with multiple liver metastases in March 1992. The patient was treated with systemic chemotherapy of 5'-deoxy-5-fluorouridine and cisplatin. After 4 courses of chemotherapy, the primary lesion responded and the multiple liver metastases disappeared. Therefore, total gastrectomy with perigastric lymphadenectomy and splenic preservation was performed. Two additional courses of the same chemotherapy were administered after the surgery. The patient has been alive without recurrence for 10 years after the curative resection.     

Role of polymorphic adhesion molecules in the development of graft-versus-leukemia effect after HLA-matched allogeneic stem cell transplantation

Mismatches of minor histocompatibility antigens (mHas) between HLA-identical stem cell donors and recipients are known as a major risk factor for graft-versus-host disease (GVHD). We determined the alleles of 5 polymorphic molecules including HA-1 and 4 adhesion molecules in 102 patients who had undergone stem cell transplantation from HLA-identical donors and investigated the association of their mismatches with the relapse rate and incidence of GVHD. We observed relapse rates of 16.1% in patients with at least one or more incompatibilities and 39.4% in patients without incompatibilities (p = 0.018). The respective relapse rates of patients with CD62L, HA-1, CD31 exon 563, CD31 exon 125 and 49b incompatibilities were 6.1%, 14.3%, 11.7%, 20% and 40%. Only patients with CD62L incompatibilities showed a lower relapse rate than patients who received a compatible graft. Since there was no difference between patients with and without incompatibilities as far as the appearance of acute GVHD (> or = 2) was concerned, we conclude that the polymorphic CD62L molecule contributes to graft-versus-leukemia rather than the development of GVHD after HLA-identical stem cell transplantation.     

Comparison of the effects of prophylactic antibiotic therapy and cost-effectiveness between cefazolin (CEZ) and Sulbactam/Ampicillin (SBT/ABPC) in gastric cancer surgery employing clinical pathway

The present study was designed to investigate the effects of prophylactic antibiotic therapy and the cost-effectiveness of Cefazolin (CEZ) and Sulbactam/Ampicillin (SBT/ABPC) in gastric cancer surgery employing clinical pathway. 157 patients (62 in the CEZ group and 95 in the SBT/ABPC group), who underwent surgery for gastric cancer at the First Department of Surgery of our hospital, were investigated. There was no significant difference between the groups with regard to sex, age, incidence of complication, stage of cancer, surgical method, operative time and blood loss, length of hospitalization, the appearance of systemic inflammatory response syndrome (SIRS), changes body temperature, white blood cell count (WBC), C-reactive protein (CRP), or clinical outcome of postoperative care by a nurse during post-operation for 7 days. The prophylactic effect of infection was also no different between the CEZ (69.4%) and SBT/ABPC (69.5%) groups. In contrast, decision analysis strongly indicated that the anticipate cost of antibiotics was higher in the latter group (yen 20402) than in the CEZ group (yen 15556), suggesting that the prophylactic effect of CEZ may be more cost-effective. Thus, evaluations of pharmacotherapy from the aspect of cost may be one of the important responsibility of hospital pharmacists in the future.     

Diagnosing dry eye using a blue-free barrier filter

PURPOSE: To investigate the benefit of the blue-free barrier filter (BF filter) in diagnosing dry eye. DESIGN: Nonrandomized clinical comparison. METHODS: Fifty-three eyes with Sj?gren syndrome (n = 42) or keratoconjunctivitis sicca (n = 11) were enrolled. Fluorescein staining was performed in temporal and nasal conjunctiva, and staining was scored with (BF score) and without the filter (FL score) from 0 to 3. Rose bengal staining was also scored (RB score) similarly. RESULTS: With FL scores of 0, 1, or 2, the BF score was significantly higher: 0.85 +/- 0.37 (P =.031), 1.74 +/- 0.65 (P =.001), and 2.65 +/- 0.48 (P =.001), respectively; with a FL score of 3, the BF score was 3 at all sites. The BF score was superior to the FL score in 66.6% of cases when the FL score was 0 and 1 and in 65.2% when the FL score was 2, but not when the FL score was 3. When the RB score was 0, 1, or 2, the BF score was higher: 1.88 +/- 0.92, 1.78 +/- 0.79, and 2.57 +/- 0.50, respectively (P =.001 for all); when the RB score was 3, the BF score was 2.97 +/- 0.16. The BF score was superior to the RB score in 75.5% of cases when the RB score was 0 and 1 and in 57.8% when the FL score was 2, but not in cases with a RB score of 3. CONCLUSIONS: The BF filter detects damaged conjunctival epithelium stained with fluorescein. Using the filter was beneficial in mild-to-moderate cases, not in severe cases. The BF filter allows diagnosis of dry eye even at the initial stage that is undetectable by conventional observation.     

Noradrenaline-induced contraction mediated by endothelial COX-1 metabolites in the rat coronary artery

Noradrenaline-induced contraction of the rat coronary arteries was significantly augmented by the presence of NG-nitro-L-arginine methyl ester (L-NAME) and arachidonic acid. The experiments in the study presented here were undertaken to characterize pharmacologically the augmented noradrenaline-induced contraction in ring preparations of rat coronary arteries. The contraction was stopped by a chemical remover of endothelium (saponin). Oxygen radical scavengers, superoxide dismutase and catalase, significantly attenuated the contraction. Cyclooxygenase-1 inhibitors (flurbiprofen, 10(-7) M) attenuated the noradrenaline-induced contraction and cyclooxygenase-2 (nimesulide, 10(-7) M) slightly attenuated the contraction. A thromboxane A2 (TXA2) synthetase inhibitor (OKY-046) and a TXA2 receptor antagonist (S-1452) did not affect the contraction. Based on these results, it was suggested that the contraction induced by noradrenaline in the rat coronary artery in the presence of L-NAME and arachidonic acid is endothelium-dependent, and that it involves reactive oxygen species and endothelial cyclooxygenase-1 metabolites of arachidonic acid.     

Effect of five-year supplementation of vitamin C on serum vitamin C concentration and consumption of vegetables and fruits in middle-aged Japanese: a randomized controlled trial

OBJECTIVE: This study was aimed at evaluating the effect of long-term vitamin C supplementation on serum and dietary vitamin C and identifying the factors associated with change in serum concentration. METHODS: A total of 439 subjects with atrophic gastritis initially participated in a randomized clinical trial using vitamin C and beta-carotene to prevent gastric cancer. We originally randomized the participants into four treatment groups using a 2x2 factorial design, whereby 0 or 15 mg/day beta-carotene and 50 or 500 mg/day vitamin C were administered in a double-blind manner. The beta-carotene component was terminated early after a mean treatment duration of four months. Before and upon early termination of beta-carotene supplementation, 134 subjects dropped out this trial, while 120 and 124 subjects took the vitamin C supplement at either 50 mg or 500 mg daily for five years. RESULTS: Changes in serum vitamin C were significantly higher in the high-dose group (38.5% increase, 95% CI = 27.0-49.9) than in the low-dose group (13.0% increase, 5.1-20.9) or in the dropout group (3.3% increase, -2.1-8.6) after five-year supplementation. The serum vitamin C at baseline was negatively associated with changes in serum vitamin C (p < 0.0001), while high-dose (p < 0.0001) and low-dose (p < 0.05) supplementation and female gender (p < 0.001) were positively associated. Dietary intake of vitamin C in the supplementation group was almost identical before and after five-year supplementation of vitamin C (2.31 mg/day decrease, 95% CI = -15.3-10.7), while a 17.7 mg/day decrease (95% CI = -44.2-8.86) was observed in the drop-out group. CONCLUSION: Five-year vitamin C supplementation induces a remarkable increase in serum vitamin C concentration, and our intervention program appears to have no effect on dietary vitamin C intake.