Membrane Fas Ligand Kills Human Peripheral Blood T Lymphocytes, and Soluble Fas Ligand Blocks the Killing

It has been believed that the Fas expressed on human peripheral blood T cells (PBT) is nonfunctional, because these cells are insensitive to agonistic anti-Fas/Apo-1 mAbs that efficiently kill in vitro–activated T cells and many Fas-expressing cell lines. Here, we demonstrate that membrane-bound Fas ligand (FasL) kills both fresh and in vitro–activated PBT, indicating that the Fas expressed on fresh PBT is functional. In contrast, soluble FasL kills only the latter. Naive T cells in umbilical cord blood do not express Fas, but can be induced to express Fas by IFN-γ or by a combination of IL-2 and anti-CD28 mAb, after which they acquire sensitivity to membrane but not to soluble FasL. Soluble FasL inhibited the killing of fresh PBT by membrane FasL. These results indicate that the shedding of FasL from the membrane is a mechanism for downregulating at least part of its killing activity.     

Characteristics of Bile Duct Carcinoma with Superficial Extension in the Epithelium

Background  Longitudinal tumor extension from the main tumor involves intramural or superficial spread along the bile duct, which influences surgical curability. Identifying the range of superficial extension is difficult by preoperative imaging. To clarify specific characteristics of bile duct carcinoma (BDC) with superficial extension of epithelium in the bile duct, we examined clinicopathologic features and patient outcomes in BDC patients with or without superficial extension who underwent surgical resection. Methods  Between 1994 and 2008, we retrospectively examined clinicopathologic findings and outcomes for 42 BDC patients who underwent surgical resection and divided them into two groups: (1) superficial extension (SE) group (n = 10); and (2) non-SE group (n = 32). Results  In terms of macroscopic growth of the main tumor, the papillary type was more common in the SE group than in the non-SE group, whereas the nodular type was dominant in the non-SE group. The prevalence of cancer-positive findings at the cut end of the bile duct was higher in the SE group. Portal vein invasion was not observed in the SE group, and the prevalence of regional lymph node metastasis was significantly greater in the non-SE group than in the SE group. No patients died of cancer in the SE group, who tended to show better survival than the non-SE group. Conclusions  The present results suggest that a good prognosis may be achieved in BDC patients with SE when complete resection is accomplished.     

Relationship Between Microvessel Count and Postoperative Survival in Patients with Intrahepatic Cholangiocarcinoma

Background  The present study aimed to elucidate the relationship between microvessel count (MVC) according to CD34 expression and prognosis in intrahepatic cholangiocarcinoma (ICC) patients who underwent hepatectomy based on our preliminary study. Methods  Relationships between MVC and clinicopathological factors were examined in 37 ICC patients. CD34 expression was analyzed using immunohistochemical methods. Results  Median MVC for ICC patients was 140/mm², which was applied as a cutoff value. Lower MVC was significantly associated with larger tumor size, periductal infiltrating type, and advanced Japanese tumor–node–metastasis stage (p < 0.05). Univariate survival analysis identified higher carcinoembryonic antigen level, periductal infiltrating type, poor histological differentiation, and lower MVC as significantly associated with lower 5-year survival rates. The 5-year survival rate in the higher-MVC group was significantly greater than that in the lower-MVC group (44% vs. 7%, p = 0.048). According to Cox multivariate survival analysis, only periductal infiltrating type on macroscopic examination was identified as a significant independent risk factor for poor survival after hepatectomy (risk ratio 4.8; p = 0.006), not MVC (1.1; p = 0.82). Conclusion  Tumor MVC might offer a useful prognostic marker of ICC patient survival after hepatectomy and further investigation in a larger series is warranted.     

Effects of methylphenidate hydrochloride on the cardiovascular system in vivo and in vitro: A safety pharmacology study

Introduction:We examined the effects of methylphenidate hydrochloride (MPH) on the cardiovascular system using in vivo and in vitro study methods in accordance with the ICH-S7B guideline.MethodsMPH was orally administered at doses of 3, 10 and 30 mg/kg to unrestrained conscious dogs implanted with a telemetry transmitter and attached with body surface electrodes, and electrocardiogram (ECG) leads. The QTcF interval was determined while heart rate (HR), and blood pressure (BP) were measured. Action potentials in isolated guinea-pig papillary muscle and the rapid component of the delayed rectifier potassium current (I_Kr) in HEK-293 cells stably transfected with hERG were also investigated at concentrations of 0.1, 0.3 and 1 µg/mL (0.37, 1.1 and 3.7 µmol/L) of MPH.ResultsNo ECG changes were observed except for a shortening of the QT interval due to a shortening of the RR interval at the maximum dose tested, 30 mg/kg. The only observed change was an elevation of BP in dogs at the dose of 30 mg/kg, which is approximately 10 times higher than the maximum therapeutic dose for use in children with attention deficit hyperactivity disorder (ADHD). Neither APD prolongation nor I_Kr inhibition was observed by MPH in the in vitro studies up to the maximum concentration tested, 1 µg/mL (3.7 µmol/L), which is approximately 34 times higher than the clinically attainable unbound plasma MPH concentrations in children with ADHD.DiscussionThese results suggest that it is unlikely that MPH affects ventricular repolarization processes at the therapeutically recommended dose levels in patients with ADHD.     

Comparative study of anastomosis in pancreaticogastrostomy and pancreaticojejunostomy after pancreaticoduodenectomy

BACKGROUND/AIMS: The relative advantages of pancreaticojejunostomy and pancreaticogastrostomy after pancreaticoduodenectomy remain to be established. To clarify differences between the two procedures, we examined 66 patients who underwent PD and compared clinical and histological parameters, surgical records and patient outcomes. METHODOLOGY: In this retrospective study, subjects were divided into a pancreaticojejunostomy group (n=48) and a pancreaticogastrostomy group (n=18). Pancreaticogastrostomy and pancreaticojejunostomy were performed using invagination technique and end-to-side anastomosis, respectively. RESULTS: There were no significant differences in patient demographics and surgical records between the two groups. Increases in serum amylase concentrations after operation were significantly greater in the pancreaticogastrostomy group (902 +/- 915 IU/L) than in the pancreaticojejunostomy group (326 +/- 761 IU/L) (p = 0.025). However, there were no significant differences in other postoperative laboratory data or complications between the two groups. CONCLUSIONS: In our study, the clinical and safety data associated with pancreaticojejunostomy and pancreaticogastrostomy procedures for anastomosis after pancreaticoduodenectomy were almost similar. Therefore, the anastomotic procedure used should depend on the surgeon's choice or the distance between the remnant stomach and pancreas.     

Two cases of epidural anesthesia using ultrasound imaging

We report two cases of epidural anesthesia in an 11-year-old obese girl (BMI 34.5) using ultrasound imaging by an inexperienced resident. It was useful for performing epidural anesthesia in these difficult cases to observe anatomical structure around epidural space and to measure the angle and distance from the skin to the epidural space. The distances in ultrasound imaging and actual puncture in case 1 were 171 mm and 250 mm, and in case 2 these were 439 mm and 480 mm, respectively. In both cases, epidural puncture was performed smoothly at the first attempt and there was no adverse effect. We conclude that using ultrasound imaging before epidural puncture in obese children is safer and more educational for residents.     

Protective targeting of high mobility group box chromosomal protein 1 in a spontaneous arthritis model

Objective

High mobility group box chromosomal protein 1 (HMGB‐1) is a DNA binding nuclear protein that can be released from dying cells and activated myeloid cells. Extracellularly, HMGB‐1 promotes inflammation. Clinical and experimental studies demonstrate that HMGB‐1 is a pathogenic factor in chronic arthritis. Mice with combined gene deficiency for DNase II and IFNRI spontaneously develop chronic, destructive polyarthritis with many features shared with rheumatoid arthritis. DNase II is needed for macrophage degradation of engulfed DNA. The aim of this study was to evaluate a potential pathogenic role of HMGB‐1 in this novel murine model.

Methods

The course of arthritis, assessed by clinical scoring and histology, was studied in DNase II^−/− × IFNRI^−/− mice, in comparison with heterozygous and wild‐type mice. Synovial HMGB‐1 expression was analyzed by immunohistochemistry. Serum levels of HMGB‐1 were determined by Western immunoblotting and enzyme‐linked immunosorbent assay (ELISA), and anti–HMGB‐1 autoantibodies were detected by ELISA. Macrophage activation was studied by immunostaining for intracellular interleukin‐1β and HMGB‐1. HMGB‐1 was targeted with truncated HMGB‐1–derived BoxA protein, acting as a competitive antagonist, with intraperitoneal injections every second day for 5 weeks.

Results

DNase II^−/− × IFNRI^−/− mice developed symmetric polyarthritis with strong aberrant cytosolic and extracellular HMGB‐1 expression in synovial tissue, in contrast to that observed in control animals. Increased serum levels of HMGB‐1 and HMGB‐1 autoantibodies were recorded in DNase II^−/− × IFNRI^−/− mice, both prior to and during the establishment of disease. Systemic HMGB‐1–specific blockade significantly ameliorated the clinical disease course, and a protective effect on joint destruction was demonstrated by histologic evaluation.

Conclusion

HMGB‐1 is involved in the pathogenesis of this spontaneous polyarthritis, and intervention with an HMGB‐1 antagonist can mediate beneficial effects.

     

Previous Laparotomy is Not a Contraindication to Laparoscopy-assisted Gastrectomy for Early Gastric Cancer

BACKGROUND: Laparoscopic procedures have generally been considered to be contraindicated in patients with a history of laparotomy because of a high risk of enteric injury during the procedure. Laparoscopy-assisted gastrectomy (LAG) has been used increasingly in the treatment of early gastric cancer, but its indication for patients with a history of laparotomy remains unclear. The aim of the present study was to estimate whether LAG is contraindicated for the patient with a history of laparotomy (PSURG). METHODS: From January 2003 to March 2006, 139 patients with early gastric cancer underwent LAG with curative intent in our institute. Fifty were PSURG patients, and the remaining 89 patients underwent LAG without any history of laparotomy (NSURG). Operative and early postoperative outcomes were compared between the groups. RESULTS: Appendectomy and gynecological surgery were the predominant procedures performed in the PSURG group prior to undergoing LAG, involving 28 patients (56.0%) and 16 patients (32.0%), respectively. Detachment of adhesion above the umbilicus was required in 25 PSURG patients (50.0%). There was no significant difference in operative and postoperative results between the two groups, although 1 PSURG patient developed symptoms of bowel injury on the first postoperative day, probably caused during the laparoscopic procedure for dissection of a jejuno-jejunal adhesion. CONCLUSIONS: There was no difference in outcome following LAG between the PSURG and NSURG groups in the present study. The PSURG patient is not contraindicated for LAG assuming careful attention is given for all operative procedures, including port insertion and dissection of intra-abdominal adhesions.