Inadequate Bowel Cleansing Efficacy of Split-dose Polyethylene Glycol for Colonoscopy in Type 2 Diabetic Patients: A Prospective and Blinded Study.

BACKGROUND/AIMS: Split-dose polyethylene glycol (PEG) is considered a standard bowel preparation regimen for colonoscopy in the general population. However, it is not clear whether the regimen is optimal for colonoscopy in diabetic patients. The aim of this study was to compare the efficacy and tolerability of split-dose PEG for diabetic versus nondiabetic patients. METHODS: This is a single-center, prospective, investigator-blinded study. A total of 55 consecutive nondiabetic and 50 diabetic patients ingested 2 L PEG solution on the day before the procedure and then 2 L of the solution on the day of colonoscopy. The quality of bowel preparation was graded using the Ottawa scale. RESULTS: There was a significant difference in bowel preparation quality, with a worse preparation except for mid colon in diabetic group (total score: 7.06±1.69 vs. 5.54±1.97, P<0.001; right colon: 2.28±0.57 vs. 1.81±0.72, P<0.001; mid colon: 1.70±0.54 vs. 1.56±0.66, P=0.253; rectosigmoid colon: 1.70±0.76 vs. 1.14±0.62, P<0.001; fluid volume: 1.38±0.53 vs. 1.01±0.59, P=0.001). About 70% of nondiabetic patients had an adequate preparation compared with only 40% of diabetic patients (P=0.003). Diabetic group had longer cecal intubation time (6.4±3.6 vs. 4.5±2.4, P=0.002) and total procedure time (22.1±7.6 vs. 18.1±8.5, P=0.015). Compliance and adverse events were not significantly different. In diabetic group, inadequate bowel preparation had a significant association with higher fasting plasma glucose (136.9±21.8 vs. 121.8±19.4 mg/dL, P=0.016). CONCLUSIONS: Diabetic patients had a worse preparation quality and longer cecal intubation and total procedure time compared with nondiabetic patients. These data suggest that split-dose PEG preparation regimen is not sufficient for optimal bowel preparation in diabetic patients undergoing colonoscopy.

Prevalence and Risk Factors of Gastroesophageal Reflux Disease in Patients with Type 2 Diabetes Mellitus.

BACKGROUND: Gastrointestinal symptoms are common in patients with type 2 diabetes mellitus (T2DM). The prevalence of gastroesophageal reflux disease (GERD) in Korea appears to be increasing. Some studies have shown that T2DM is a risk factor for symptomatic GERD. However, this possibility is still debated, and the pathogenesis of GERD in T2DM is not yet fully understood. The aim of this study was to analyze the prevalence and risk factors (including autonomic neuropathy) of GERD in patients with T2DM. METHODS: This cross-sectional case-control study enrolled T2DM patients (n=258) and healthy controls (n=184). All participants underwent physical examinations and laboratory tests. We evaluated medical records and long-term diabetes complications, including peripheral and autonomic neuropathy in patients with T2DM. Esophagogastroduodenoscopy was performed in all patients. The Los Angeles (LA) classification was used to grade GERD. GERD was defined as LA grade A (or higher) or minimal change with GERD symptoms. GERD symptoms were examined using a frequency scale. Data were expressed as mean±standard error. Independent t-tests or chi-square tests were used to make comparisons between groups. RESULTS: The prevalence of GERD (32.6% vs. 35.9%, P=0.266) and GERD symptoms (58.8% vs. 59.2%, P=0.503) was not significantly different between T2DM patients and controls. We found no significant differences between T2DM patients with GERD and T2DM patients without GERD with respect to diabetic complications, including autonomic neuropathy, peripheral neuropathy, duration of DM, and glucose control. CONCLUSION: The prevalence of GERD in patients with T2DM showed no difference from that of controls. GERD was also not associated with peripheral and cardiovascular autonomic neuropathy, age, or duration of DM in patients with T2DM.

The Effectiveness of Short Message Service to Assure the Preparation-to-Colonoscopy Interval before Bowel Preparation for Colonoscopy.

Background/Aims. The preparation-to-colonoscopy (PC) interval is one of several important factors for the bowel preparation. Short message service (SMS) reminder from a cellular phone has been suggested to improve compliance in various medical situations. We evaluated the effectiveness of SMS reminders to assure the PC interval for colonoscopy. Methodology. This prospective randomized study was investigator blinded. In the No-SMS group, patients took the first 2 L polyethylene glycol (PEG) between 6 and 8 PM on the day before colonoscopy and the second 2 L PEG approximately 6 hours before the colonoscopy without SMS. In the SMS group, patients took first 2 L PEG in the same manner as the No-SMS group and the second 2 L PEG after receiving an SMS 6 hours before the colonoscopy. Results. The SMS group had a lower score than the No-SMS group, according to the Ottawa Bowel Preparation Scale (P < 0.001). Multivariate logistic regression analysis showed that compliance with diet instructions (odds ratio (OR) 2.109; 95% confidence interval (CI), 1.11-3.99, P = 0.022) and intervention using SMS ((OR) 2.329; 95% (CI), 1.34-4.02, P = 0.002) were the independent significant factors for satisfactory bowel preparation. Conclusions. An SMS reminder to assure PC interval improved the bowel preparation quality for colonoscopy with bowel preparation.

Comparison of the Efficacy and Tolerability between Same-day Picosulfate and Split-dose Polyethylene Glycol Bowel Preparation for Afternoon Colonoscopy: A Prospective, Randomized, Investigator-blinded Trial.

BACKGROUND/AIMS: In the present study, we evaluated the efficacy and tolerability between same-day bowel preparation protocols using 2 sachets of Picosulfate and a 4 L split-dose polyethylene glycol (PEG) bowel preparation for afternoon colonoscopy. METHODS: The study had a single-center, prospective, randomized, and investigator-blinded, non-inferiority design. We evaluated bowel preparation quality according to the Ottawa scale, patient tolerability, compliance, incidence of adverse events, sleep quality, and polyp/adenoma detection rate. RESULTS: Among the 196 patients analyzed (mean age, 55.3 years; 50.3% men), 97 received the same-day regimen of 2 sachets of picosulfate (group A) and 99 received the 4 L split-dose PEG regimen (group B). The Ottawa score of the total colon was 4.05±1.56 in group A and 3.80±1.55 in group B (P=0.255). The proportion of patients having adequate bowel preparation in the same-day picosulfate group (61.5%) was slightly less than the 4 L PEG group (71.3%); however, the difference was not statistically significant (P=0.133). Tolerability of the group A regimen was superior to that of the group B regimen (P<0.000). The same-day picosulfate regimen was associated with fewer adverse events, such as abdominal bloating (P=0.037) and better sleep quality (P<0.000). CONCLUSIONS: The same-day picosulfate regimen and the 4 L split-dose PEG regimen had similar efficacy in bowel preparation for afternoon colonoscopy. However, the same-day picosulfate regimen was easier to administer, produced fewer adverse events, and enabled better sleep quality.

The Effect of Indigocarmine on Improvement of the Polyp Detection Rate during Colonoscopic Examination with Hood Cap.

BACKGROUND/AIMS: Hood cap-assisted chromocolonoscopy using indigocarmine is expected to improve the detection rate of colorectal polyps, especially adenomatous polyps. Therefore, aim of the present study was to evaluate the usefulness of hood cap-assisted chromocolonoscopy in routine colonoscopic examinations. METHODS: From January, 2013 through March, 2013, a total of 86 patients were enrolled (M:F=33:53, mean age=60 years). For each patient, hood cap-assisted colonoscopic examination was performed, followed by hood cap-assisted chromocolonoscopy using 0.2% indigocarmine from the cecum to the hepatic flexure. Total numbers and characteristics of polyps were compared before and after indigo carmine dye spraying. RESULTS: Prior to dye spraying, 48 polyps were found in 37 patients, and after dye spraying, 53 additional polyps were found in 34 patients. Of these undetected polyps, 45 (85%) were small sized polyps (≤0.5 cm). Histologically, 19 (36%) were adenomatous polyps, and of these, 15 (28%) were tubular adenomas and 4 (8%) were serrated adenomas. As for the polyp detection rate, there was no difference between the expert and the non-expert groups. CONCLUSION: Hood cap-assisted chromocolonoscopic examination using indigocarmine was helpful in detecting cecum and ascending colon polyps, especially small sized polyps (<0.5 cm) and neoplastic polyps.

Low-volume morning-only polyethylene glycol with specially designed test meals versus standard-volume split-dose polyethylene glycol with standard diet for colonoscopy: a prospective, randomized trial.

BACKGROUND/AIMS: Split-dose polyethylene glycol (PEG) is a standard bowel preparation regimen for colonoscopy, but the large volume is burdensome to ingest and the night dose causes sleep disturbance. This study was performed to evaluate the efficacy and tolerability of a bowel preparation protocol using low-volume morning-only PEG with specially designed low-residue test meals (LV-PEG with TM) compared to a standard-volume split-dose PEG bowel preparation with a standard diet (SV-PEG with SD). METHODS: This was a single-center, prospective, randomized, investigator-blinded, noninferiority study. The primary end point was bowel preparation quality according to the Ottawa scale. Tolerability, compliance, adverse events, sleep quality and polyp/adenoma detection were also assessed. RESULTS: Among 197 patients analyzed (mean age 54.6 years, 51.3% men), 97 received LV-PEG with TM and 100 received SV-PEG with SD. The Ottawa score for the total colon was 3.76 ± 2.07 in the LV-PEG with TM group and 3.67 ± 1.57 in the SV-PEG with SD group (p = 0.723; difference 0.09, 95% confidence interval -0.60 to 0.42). The compliance was high (more than 95%) in both groups (p = 0.621). PEG was easier to ingest for patients in the LV-PEG with TM group compared to the SV-PEG with SD group [visual analogue scale (VAS) for difficulty: 4.64 ± 2.46 vs. 5.97 ± 2.42, respectively; p < 0.001]. Diet instructions were also easier to comply with for patients in the LV-PEG with TM group compared to the SV-PEG with SD group (VAS for difficulty: 3.11 ± 2.25 vs. 4.00 ± 2.39, respectively; p = 0.008). Patients in the LV-PEG with TM group had a lower incidence of abdominal bloating (p = 0.012) and better sleep quality (p < 0.001). There was no difference between the groups regarding polyp and adenoma detection. CONCLUSIONS: LV-PEG with TM and SV-PEG with SD have similar efficacy with regard to bowel preparation for colonoscopy. LV-PEG with TM provided easier PEG intake and diet compliance, less abdominal bloating and better sleep quality than SV-PEG with SD.

Determination of the optimal time for premedication with pronase, dimethylpolysiloxane, and sodium bicarbonate for upper gastrointestinal endoscopy.

GOALS AND BACKGROUND: Premedication with pronase, dimethylpolysiloxane, and sodium bicarbonate improves visibility during upper gastrointestinal (UGI) endoscopy. However, the optimal time for this combination to take effect is unknown. We investigated the optimal time of pre-UGI endoscopy medication. STUDY: A randomized, investigator-blind, controlled trial. The 300 patients who were to receive premedication were randomized into 3 groups according to the following medication time before UGI endoscopy: 10 minutes (group A, n=98), 10 to 30 minutes (group B, n=97), and 30 minutes premedication (group C, n=99). Visibility scores (range, 1 to 4, with lower scores indicating better gastric mucosal visibility) were assessed for the antrum, lower body, upper body, and fundus and compared, including the sum of the scores, between the 3 groups. RESULTS: Group B had significantly lower visibility scores for the lower body, upper body, and fundus than group C (P=0.001, 0.009, and 0.002, respectively). Group A obtained significantly lower scores for the antrum and lower body than group C (P=0.007 and 0.005, respectively). The total visibility scores of groups A and B were significantly lower compared with those of group C (P=0.001, 0.003, respectively). CONCLUSIONS: Administration of pronase, dimethylpolysiloxane, and sodium bicarbonate within 30 minutes before UGI endoscopy significantly improved endoscopic visualization. However, the optimal time to achieve the best visibility was between 10 to 30 minutes before UGI endoscopy.

The efficacy and safety of carbon dioxide insufflation during colonoscopy with consecutive esophagogastroduodenoscopy in moderately sedated outpatients: a randomized, double-blind, controlled trial.

GOALS AND BACKGROUND: Colonoscopy with consecutive esophagogastroduodenoscopy (CCEGD) can be more convenient than performing each procedure individually. There has been no randomized controlled trial comparing carbon dioxide (CO2) versus air insufflations during CCEGD in sedated patients. CO2 insufflation instead of air during CCEGD may reduce abdominal pain and be more comfortable. We investigated the efficacy and safety of CO2 insufflation during CCEGD in moderately sedated outpatients. STUDY: This was a randomized, double-blind, controlled trial. A total of 96 outpatients were randomly assigned to the groups of CO2 or air insufflation. Postprocedure pain was assessed using a 0 to 10 visual analogue scale, and the proportion of pain-free patients was compared between the groups. Waist circumferences and end-tidal CO2 (ETCO2) were measured. RESULTS: Among 96 patients, cecal intubation failed in 2 patients, who were excluded from the analysis. Forty-eight patients in the CO2 and 46 patients in the air group completed the study. There was significant difference between the 2 groups regarding the proportion of pain-free patients 30 minutes after the procedures [air group, 35/46 (76.1%) vs. CO2 group, 44/48 (91.6%)] (P=0.03). However, there was no significant difference in the proportion at 6 and 24 hours after the procedures. The mean increase in waist circumference was greater with air than with CO2 (1.54 vs. 0.18 cm, P<0.001). The ETCO2 measured immediately after the procedures was slightly higher in the CO2 group than in the air group (38.6 vs. 37.2 mm Hg, P=0.02), but the values were within the normal range. No significant adverse events occurred. CONCLUSIONS: CO2 insufflation during CCEGD reduced postprocedural pain and distension compared with air. It was comfortable and safe to use in moderately sedated outpatients.

Efficacy of prokinetics with a split-dose of polyethylene glycol in bowel preparation for morning colonoscopy: a randomized controlled trial.

BACKGROUND/AIMS: Currently, a split-dose of polyethylene glycol (PEG) is the mainstay of bowel preparation due to its tolerability, bowel-cleansing action, and safety. However, bowel preparation with PEG is suboptimal because residual fluid reduces the polyp detection rate and requires a more thorough colon inspection. The aim of our study was to demonstrate the efficacy of a sufficient dose of prokinetics on bowel cleansing together with split-dose PEG. METHODS: A prospective endoscopist-blinded study was conducted. Patients were randomly allocated to two groups: prokinetic with split-dose PEG or split-dose PEG alone. A prokinetic [100 mg itopride (Itomed)], was administered twice simultaneously with each split-dose of PEG. Bowel-cleansing efficacy was measured by endoscopists using the Ottawa scale and the segmental fluidity scale score. Each participant completed a bowel preparation survey. Mean scores from the Ottawa scale, segmental fluid scale, and rate of poor preparation were compared between both groups. RESULTS: Patients in the prokinetics with split-dose PEG group showed significantly lower total Ottawa and segmental fluid scores compared with patients in the split-dose of PEG alone group. CONCLUSION: A sufficient dose of prokinetics with a split-dose of PEG showed efficacy in bowel cleansing for morning colonoscopy, largely due to the reduction in colonic fluid.

Optimal preparation-to-colonoscopy interval in split-dose PEG bowel preparation determines satisfactory bowel preparation quality: an observational prospective study.

BACKGROUND: Several factors influence bowel preparation quality. Recent studies have indicated that the time interval between bowel preparation and the start of colonoscopy is also important in determining bowel preparation quality. OBJECTIVE: To evaluate the influence of the preparation-to-colonoscopy (PC) interval (the interval of time between the last polyethylene glycol dose ingestion and the start of the colonoscopy) on bowel preparation quality in the split-dose method for colonoscopy. DESIGN: Prospective observational study. SETTING: University medical center. PATIENTS: A total of 366 consecutive outpatients undergoing colonoscopy. INTERVENTION: Split-dose bowel preparation and colonoscopy. MAIN OUTCOME MEASUREMENTS: The quality of bowel preparation was assessed by using the Ottawa Bowel Preparation Scale according to the PC interval, and other factors that might influence bowel preparation quality were analyzed. RESULTS: Colonoscopies with a PC interval of 3 to 5 hours had the best bowel preparation quality score in the whole, right, mid, and rectosigmoid colon according to the Ottawa Bowel Preparation Scale. In multivariate analysis, the PC interval (odds ratio [OR] 1.85; 95% CI, 1.18-2.86), the amount of PEG ingested (OR 4.34; 95% CI, 1.08-16.66), and compliance with diet instructions (OR 2.22l 95% CI, 1.33-3.70) were significant contributors to satisfactory bowel preparation. LIMITATIONS: Nonrandomized controlled, single-center trial. CONCLUSIONS: The optimal time interval between the last dose of the agent and the start of colonoscopy is one of the important factors to determine satisfactory bowel preparation quality in split-dose polyethylene glycol bowel preparation.

Efficacy and tolerability of split-dose PEG compared with split-dose aqueous sodium phosphate for outpatient colonoscopy: a randomized, controlled trial.

BACKGROUND AND AIMS: Adequate bowel cleansing is essential for a high-quality, effective, and safe colonoscopy. The aims of this study were to compare the efficacy and tolerability of split-dose polyethylene glycol with aqueous sodium phosphate for outpatients who underwent scheduled colonoscopy. METHODS: In this prospective trial, 207 outpatients (aged between 18 and 65 years, with normal renal function, at low risk for renal damage) were randomized to receive split-dose preparation of PEG (2 L/2 L) (N = 103) or NaP (45 mL/45 mL, 12 h apart) (N = 104) without strict diet restriction the day before colonoscopy. The bowel cleansing efficacy of preparations was rated according to the Ottawa scale and the patient tolerability was assessed using a patient questionnaire. RESULTS: There was no significant difference between the two groups for the mean total score using the Ottawa bowel preparation scale (P = 0.181). Significantly greater residual colonic fluid was observed in the split-dose PEG group (1.24 ± 0.49) than in the NaP group (1.04 ± 0.53) (P = 0.007). Patient compliance, preference, and acceptance of a two preparation regimen were similar with no significant differences (P = 0.095, P = 0.280 and P = 0.408, respectively). The overall incidence of adverse events was not significantly different between the two groups; however, the split-dose PEG group tended to have fewer adverse events (52/103 [50.5%], 66/104 [63.5%], P = 0.059) and had significantly less nausea and vomiting (P = 0.036). CONCLUSIONS: Split-dose PEG, compared with split-dose NaP, is associated with more residual colonic fluid, but produces equivalent colon cleansing efficacy and results in less nausea and vomiting, which might improve patient tolerability (clinical trial registration number NCT01229800).

The efficacy of the combination therapy with oral and topical mesalazine for patients with the first episode of radiation proctitis.

BACKGROUND/AIMS: Radiation proctitis is a common complication of pelvic radiation for which an optimal treatment remains undetermined. We assessed the efficacy of oral and topical mesalazine combination therapy for patients with naive radiation proctitis. METHODS: A total of 23 patients with radiation proctitis were enrolled in the study over a period of 2 years. Three of these patients were excluded due to severe bleeding during the study. Twenty patients (mean age 60.3 years; two males, 18 females) were treated with oral mesalazine (3 × 1 g per day) plus a daily mesalazine suppository (1 g per day at bedtime) for 4 weeks. The efficacy of treatment was assessed according to the Subjective Objective Management Analytic (SOMA) scale for alleviation of clinical symptoms of rectal toxicity and sigmoidoscopic findings. RESULTS: The mean bleeding score improved significantly from 2.10 to 1.70 (p = 0.002) with mesalazine treatment. However, scores were not improved for pain (0.30-0.20, p = 0.163), tenesmus (0.50-0.45, p = 0.577), or stool frequency (0.35-0.30, p = 0.577). The improvements in the mean telangiectasia score (1.80-1.45, p = 0.005), bleeding point score (1.60-1.05, p < 0.001), and friable mucosa score (1.35-1.00, p = 0.005) were all statistically significant. No side-effects were noted in any of the patients. CONCLUSIONS: The combination of oral and topical mesalazine therapy for radiation proctitis may be a safe and effective treatment for naive radiation proctitis, especially for hemorrhagic proctitis. A large, randomized controlled trial is required to confirm the results of this pilot study.

The effect of wireless LAN-based PACS device for portable imaging modalities.

The aim of this study was to develop wireless Picture Archiving and Communication System (PACS) device and to analyze its effect on image transfer from portable imaging modalities to the main PACS server. Using a laptop computer equipped with wireless local area network (LAN), the authors developed a wireless PACS device with DICOM modality worklist and DICOM storage server modules. This laptop computer could be easily fixed to portable imaging modalities such as ultrasound machines. From May to August 2007, 112 portable examinations were evaluated. Of these, 62 were done with wireless LAN-based PACS device, and 50 were done without wireless PACS device. To evaluate the impact of the wireless LAN-based PACS device on productivity and workflow, we analyzed the mean time delay and standard deviations (SD) both in cases where wireless LAN-based PACS device was used and in cases where it was not used. Statistical analysis was performed using a t test. The mean time interval from image acquisition to storage in the main PACS when the wireless LAN-based PACS device was used was 342.4 s (5 min and 42.4 s, SD = 509.2 s). When the wireless PACS was not used, the mean time interval was 2,305.5 s (38 min and 25.5 s, SD = 1,371.8 s). The mean time interval was statistically different between the two groups (t test, p < 0.001). The wireless LAN-based PACS device could help in reducing the storage intervals of images obtained by portable machines and in promoting effective and rapid treatment of patients who have undergone portable imaging examinations.

Detection of expressed IL-32 in human stomach cancer using ELISA and immunostaining.

Interleukin (IL)-32 is a recently identified proinflammatory cytokine that is one of the IL-18 inducible genes, and plays an important role in autoimmune and inflammatory diseases. We produced antibodies against IL-32 and studied the expression of IL-32 in human stomach cancer. We detected IL-32 secreted from K-562 cells that werw stably transfected with IL-32 and in the sera of stomach cancer patients, by a sandwich ELISA using a monoclonal antibody KU32-52 and a polyclonal antibody. In order to optimize a sandwich immunoassay, recombinant IL-32alpha was added, followed by the addition of a biotinylated KU32-52 into microtiter plate wells precoated with a goat anti-IL-32 antibody. The bound biotinylated KU32-52 was probed with a streptavidin conjugated to HRP. This sandwich ELISA was highly specific and had a minimal detection limit of 80 pg/ml (mean+/-SD of zero calibrator) and measuring up to 3,000 pg/ml. This ELISA showed no cross-reaction with other cytokines such as hIL-1alpha, hIL-1beta, hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-alpha. Intra-assay coefficients of variation were 18.5% to 4.6% (n=10), and inter-assay coefficients were 23% to 9% (n=10). The average IL-32 level in the sera of 16 stomach cancer patients (189 pg/ml) was higher than that of 12 healthy control men (109 pg/ml). Our results indicate that serum IL-32 level can be detected by using an established ELISA, and that this immunoassay and mAb KU32-09 specific for immunohistochemistry can be used in the detection of expressed and secreted IL- 32 in stomach cancer patients.

[Comparison of transient elastography and hepatic fibrosis assessed by histology in chronic liver disease].

BACKGROUND/AIMS: Transient elastography (Fibroscan, Echosens, France) is a new, simple, and non-invasive method to assess the degree of hepatic fibrosis by measuring liver stiffness. Recent reports have shown that liver stiffness measurement using Fibroscan allowed accurate prediction of hepatic fibrosis in patient with chronic hepatitis C. The aim of this study was to evaluate accuracy of Fibroscan for the detection of hepatic fibrosis in Korea with various etiologies of chronic liver disease by comparison with fibrosis assessed by histologic examination. METHODS: Fifty-four patients with chronic liver diseases, which were histologically confirmed within recent 6 months were enrolled. Etiologies were HBV, HCV infection, autoimmune hepatitis, and non alcoholic steatohepatitis. Hepatic fibrosis was graded on the basis of standard guideline proposed by the Korean Study Group for the Pathology of Digestive Diseases. RESULTS: Fibroscan values were significantly higher in F3 (16.96 kPa) and F4 (19.86 kPa) than others (p=0.003). Liver stiffness measurement was significantly correlated to the fibrosis stage (r=0.614, p<0.0001). CONCLUSIONS: Liver stiffness measurement by Fibroscan is a promising method for the assessment of hepatic fibrosis in chronic liver disease because it accompanies no complication.

KR-31831, benzopyran derivative, inhibits VEGF-induced angiogenesis of HUVECs through suppressing KDR expression.

Angiogenesis is important in the development and progression of cancer, therefore the therapeutic approach based on anti-angiogenesis may represent a promising therapeutic option. KR-31831 is a novel anti-ischemic agent. Previously, we reported the anti-angiogenic activity of KR-31831. In the present study we investigated the molecular mechanisms underlying anti-angiogenic activity of KR-31831. We show that KR-31831 inhibits vascular endothelial growth factor (VEGF)-induced proliferation and tube formation via release of intracellular Ca2+ and phosphorylation of extra-cellular regulated kinase 1/2 (Erk 1/2) in human umbilical vein endothelial cells (HUVECs). Moreover, the expression of VEGF receptor 2 (VEGFR2, known as Flk-1 or KDR) was reduced by the treatment of KR-31831. These results suggest that KR-31831 may have inhibitory effects on tumor angiogenesis through down-regulation of KDR expression.

Effects of a tetramethoxyhydroxyflavone p7f on the expression of inflammatory mediators in LPS-treated human synovial fibroblast and macrophage cells.

The inhibitory effects of 5,6,3',5'-tetramethoxy 7,4'-hydroxyflavone (labeled as p7F) were elucidated on the productions of proinflammatory cytokines as well as inflammatory mediators in human synovial fibroblasts and macrophage cells. p7F inhibited IL-1beta or TNF-alpha induced expressions of inflammatory mediators (ICAM-1, COX-2, and iNOS). p7F also inhibited LPS-induced productions of nitric oxide and prostaglandin E2 in RAW 264.7 cells. In order to investigate whether p7F would inhibit IL-1 signaling, p7F was added to the D10S Th2 cell line (which is responsive to only IL-1beta and thus proliferates), revealing that p7F inhibited IL-1beta-induced proliferation of D10S Th2 cells in a doseresponse manner. A flow cytometric analysis revealed that p7F reduced the intracellular level of free radical oxygen species in RAW 264.7 cells treated with hydrogen peroxide. p7F inhibited IkappaB degradation and NF-kappaB activation in macrophage cells treated with LPS, supporting that p7F could inhibit signaling mediated via toll-like receptor. Taken together, p7F has inhibitory effects on LPS-induced productions of inflammatory mediators on human synovial fibroblasts and macrophage cells and thus has the potential to be an antiinflammatory agent for inhibiting inflammatory responses.

Interleukin-32 monoclonal antibodies for immunohistochemistry, Western blotting, and ELISA.

The members of the IL-1 family play important roles in the development and pathogenesis of autoimmune and inflammatory diseases. Especially, IL-1 and IL-18 belong to the IL-1 family because they share structural similarity and require caspase-1 for processing. Currently, IL-18 has been studied for its biological effects in the broad spectrum of Th1- or Th2- related autoimmune diseases. IL-18 also uses a similar signaling pathway as that of IL-1 family members. Taken together these results, IL-18-inducible genes might also contribute to autoimmune and inflammatory diseases. It has recently been reported that an inducer of TNF-alpha was identified as one of IL-18 inducible genes in IL-18 responsible cells and named as a new cytokine IL-32. We have produced novel monoclonal anti IL-32 antibodies in order to help study IL-32 function and to develop improved diagnosis of IL-32-expressing tumors. Several mAbs reactive to IL-32 isoforms were prepared and characterized by the epitope analysis and Western blotting performed using various deletion mutants and IL-32 isoforms (IL-32alpha, beta, gamma, and delta). In order to optimize the sandwich ELISA for IL-32, recombinant IL-32alpha was added, followed by the addition of a biotinylated mAb KU32-52 into the microtiter plate wells pre-coated with a mAb KU32-07 or mAb KU32-56. The bound mAb was probed with a streptavidin conjugated to HRP. The epitope analysis and Western blot analysis revealed that mAb KU32-07 could detect only IL-32alpha and KU32-52 was bound to all isoforms, whereas KU32-56 were reactive to IL-32 alpha, beta, delta isoforms but not gamma isoform. These sandwich ELISAs were highly specific and had a minimal detection limit of 80 pg/ml (mean+3 SD of zero calibrator) and measuring range of up to 3000 pg/ml. An ELISA using a coating mAb KU32-07 and a capturing biotinylated mAb KU32-52 had no cross-reaction with other cytokines such as IL-32beta, IL-32gamma, IL-32delta, hIL-1alpha , IL-1beta , hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-alpha. Intra-assay coefficients of variation were 11 to 6% (n=16) and inter-assay coefficients were 10 to 5% (n=9). Another ELISA using a coating mAb KU32-56 and a capturing biotinylated mAb KU32-52 detected both IL-32alpha and IL-32beta isoforms but not gamma and delta isoforms and had no cross-reaction with other cytokines such as hIL-1alpha , IL-1beta , hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-alpha. One mAb KU32-09 was shown to react strongly on immunohistochemistry. Our newly established mAbs, KU32-07, KU32-09, KU32-52, KU32-56, have different and useful properties for the detection of IL-32 by immunohistochemistry, ELISA, and Western blotting.

Down-regulation of UNC5 homologue expression after the spinal cord injury in the adult rat.

UNC5A-D are transmembrane receptors for netrin that plays an important role in the development of the spinal cord. However, their function in the adult nervous system remains to be established. In the present study, the mRNA expression of UNC5 family was investigated in the adult rat spinal cord following a dorsal hemisection injury. Reverse transcription-polymerase chain reaction (RT-PCR) revealed that mRNA expression of rat UNC5 family was down-regulated in the spinal cord within 3 days after the dorsal hemisection. Unc5b-d mRNA expressions returned to near normal level after a week, whereas the down-regulation of Unc5a mRNA expression was maintained until 1 month after the injury. In situ hybridization and immunofluorescent showed that both neurons and non-neuronal cells express Unc5a and b mRNAs in the adult spinal cord, and the significant long lasting down-regulation of Unc5a mRNA expression was also noticed following the spinal cord injury. This result suggests netrin/UNC5 involvement in the injury response of the adult spinal cord of the rats.

[A case of cholestatic hepatitis induced by epstein-barr virus infection].

Acute viral hepatitis in human can be caused by a large number of viruses with a wide range of clinical manifestations and laboratory findings. EBV is a rare causative agent of an acute hepatitis, during the course of infectious mononucleosis. Hepatic manifestations of EBV are usually mild and resolve without serious complications. EBV is rather uncommonly confirmed as an etiologic agent in acute viral hepatitis of adults and it rarely causes cholestatic hepatitis. We report a case of EBV hepatitis with cholestatic feature that was verified through serum viral marker and liver biopsy.