Accumulation of 1-methyl-tetrahydro-Β-carboline-3-carboxylic acid in blood and organs of rat. A possible causative substance of eosinophilia-myalgia syndrome associated with ingestion ofl-tryptophan

1-Methyl-tetrahydro--carboline-3-carboxylic acid (MTCA) may cause eosinophilia-myalgia syndrome (EMS) associated with ingestion ofl-tryptophan. The distribution and excretion of MTCA were studied in rats which had received perorally a single 1.6 mg/kg dose of MTCA. MTCA concentrations in blood, kidney, liver, brain, heart, spleen, lung and gastrocnemius muscle were measured by HPLC combined with fluorometric detection. The concentration of MTCA in each organ reached a maximum at 1 h and then gradually declined. However, a significant level of MTCA still remained at 5 h, when 52% of ingested MTCA remained in the contents of the large intestine. Twenty-nine percent of the ingested MTCA was excreted in urine over the course of 24 h. A higher dose (10 mg/kg) of MTCA resulted in significant elevations in the concentrations and amounts of MTCA in the various organs. In addition, chronic treatment with a 10 mg/kg dose of MTCA for 6 weeks further increased the concentrations and amounts of MTCA in each organ. However, no histological changes were observed in any of the organs after chronic treatment. This is the first report which demonstrates accumulation of MTCA in the blood and various organs, including muscle, of rats.     

Growth suppression of non-small cell lung carcinoma cells by the introduction of the p16(INK4A) gene

The p16(INK4A) gene is frequently inactivated in nonsmall cell lung carcinoma (NSCLC) by either mutations, deletions or DNA methylations. To assess the biological significance of p16(INK4A) inactivation in the development of NSCLC, full-length p16(INK4A) cDNA was introduced into NSCLC cell lines, A549 and H322, in which p16(INK4A) was homozygously deleted. NSCLC cells transfected with the p16(INK4A) expression vector formed colonies in 20-68% of those with a control vector, and exogenous p16(INK4A) protein was expressed in 4 of 68 A549-derived clones and none of 29 H322-derived clones, respectively. A549-derived clones which stably expressed the exogenous p16(INK4A) gene showed significant decrease in growth rate in vitro and tumorigenicity in vivo in proportion to the level of p16(INK4A) expression. Furthermore, the cell cycle of these cells significantly delayed with accumulation of cells in G1 phase. Micro-injection of p16(INK4A) expression vector also revealed that p16(INK4A) blocked S phase entry in both A549 and H322 cells. These results suggest that the restoration of the p16(INK4A) function suppresses the growth of NSCLC cells by induction of G1 arrest in the cells. Therefore, inactivation of p16(INK4A) may play an important role in the enhancement of unregulated NSCLC growth in vivo.     

Evaluation of esophageal bile reflux after total gastrectomy by gastrointestinal and hepatobiliary dual scintigraphy

Conducting the qualitative evaluation of reconstruction methods is difficult because of their complexity. The aim of the present study was to compare esophageal bile and food reflux by performing gastrointestinal and hepatobiliary dual scintigraphy (GHDS) after various methods of reconstruction following total gastrectomy. Of 17 patients studied, 4 had undergone Roux-en-Y anastomoses (R-Y); 6, jejunal pouch-Y anastomoses (P-Y); and 7, jejunal pouch interposition (P-I). GHDS was performed 1 year after surgery using¹¹¹In-diethylene triamine pentaacetic acid administered orally, and^99mTc-pyridoxyl-5-methyl tryptophan administered intravenously. Imaging data from a gamma camera were stored in and processed by a data analyzer. Three patients who had undergone R-Y and one who had undergone P-I complained of heartburn, while one who had undergone R-Y, two who had undergone P-Y, and three who had undergone P-I complained of a feeling of fullness. Esophageal bile reflux was confirmed by GHDS in four of the patients who had undergone R-Y, one who had undergone P-Y, and four who had undergone P-I. Moreover, GHDS demonstrated food retention in two patients who had undergone R-Y, five who had undergone P-Y, and four who had undergone P-I. Weight loss was closely related to the esophageal reflux of bile or food which can be accurately detected by GHDS. Despite the absence of heartburn, patients diagnosed as having bile reflux by GHDS showed poor recovery of body weight.     

Long-term outcome of living related liver transplantation for patients with intrapulmonary shunting and strategy for complications

BACKGROUND: In 320 living related liver transplantation performed between June 1990 and September 1997, there were 21 living related liver transplantation for patients with intrapulmonary shunting, manifested by digital clubbing, cyanosis, and dyspnea. We report the long-term outcome for more 6 months and our strategy to overcome complications in these recipients. PATIENTS: A total of 21 patients (age range 2-33 years, 19 children and 2 adults, 6 males and 15 females) were classified into three grades according to shunt ratio calculated by TcMAA pulmonary scintigraphy; 5 in mild group (shunt ratio: less than 20%), 6 in moderated group (20%-40%), and 10 in severe group (more than 40%). The original underlying liver disease was biliary atresia in all patients. RESULTS: Spearmen's correlation coefficient rank test revealed that shunt ratio correlated significantly with PaO2 in room air (P=0.0001), PaO2 in 100% oxygen (P=0.0004), hematocrit (P=0.0276), and period of dyspnea before transplantation (P=0.023). COMPLICATIONS: Wound infection occurred in 80, 66, and 80%, and bile leakage in 20, 0, 40% in mild, moderate, and severe group, respectively. Patients who had portal vein thrombosis, and intracranial complication were classified as severe group and the incidence was 20 and 20%, respectively. The patient actuarial one year survival was 80, 66.7, and 48%, in mild, moderate, and severe group, respectively, although there was no significant difference. All patients who survived improved hepatopulmonary syndrome and the length of period required for the resolution was significantly correlated to the preoperative shunt ratio (P=0.023). COMMENTS: Patients with severe shunting are susceptible to wound infection and bile leak. The trend of higher incidence of portal thrombosis and intracranial complications in the severe group was closely related high hematocrit. Secure surgical technique to reduce bile leak and delayed primary wound closure to reduce wound infection were found to be effective. Anticoagulant therapy by infusing heparin through the portal vein followed by coumadin could prevent fatal portal vein thrombosis without counter risk of fatal cerebral hemorrhage.     

Present situation of medical care for foreigners at medical facilities in Chiba City]

OBJECTIVE: The number of foreigners residing in Japan has increased during the past decade. The aim of this survey was to clarify the present situation and the disadvantages in the medical care for foreigners. METHODS: In October 1997, we mailed a questionnaire to all clinics and hospitals in Chiba City. The questionnaire included questions regarding the number of foreign patients who visited during the past three months, the proportion of patients who have any health insurance, procedures for handling foreign patients, and the disadvantages in medical care for them. RESULTS: Of the 210 respondents (183 clinics and 27 hospitals), 133 facilities (63.3%) provided medical care for foreign patients during the past three months, and 187 (89.0%) during the past year. In 102 facilities (76.7%), the number of foreign patients was fewer than 10 during the past three months. The mean number of foreign patients was 13.8 per facility (12.2 per clinic, and 23.5 per hospital). Of these foreign patients, 68.6% had any health insurance. In 172 facilities (81.9%), foreign patients were accepted in the same way as the Japanese. The main disadvantages in medical care for foreigners were difficulty in understanding because of communication problems, and default in payment of medical expenses for the foreigners with no health insurance. In 154 facilities (73.3%), no special preparation to communicate with foreign patients was provided, and few facilities could understand foreign languages except for English. In 20 facilities, fees for medical care had been left unpaid by foreign patients during the past year, and the total outstanding amount was about 23,800,000 yen. CONCLUSION: In Chiba City, many medical facilities provide medical care for foreign patients. This study suggests that support for communication with foreigners and compensation for default in medical expenses are necessary.     

A study of factors influencing determination of discharge disposition of stroke rehabilitation patients]

PURPOSE: The purpose of this study is to statistically analyze medical and socioeconomic factors which influence discharge disposition from a stroke unit. METHODS: We investigated 126 stroke patients admitted to a community hospital within 30 days from the onset of the stroke. Mean age was 65.9 +/- 13.9 (SD) years old, profile of diagnosis was 89 infarctions was 37 hemorrhages, and mean length of stay was 79.7 +/- 45.1 days. Our rehabilitation team consists of doctors, nurses, physical therapists, occupational therapists and a medical social worker. Our stroke unit accepted the stroke patients from the time of admission and attempted to begin rehabilitation as soon as possible. The patients and their families were informed about their options after discharge from the hospital. Eight factors were analyzed using a multiple logistic regression model. RESULTS: 1) Ninety-eight patients (77.8%) could return home (home group) and 28 patients (22.2%) were discharged to another hospital or a long-term care facility (LTCF group). 2) Mean age of LTCF group (70.3 +/- 12.0) was significantly higher than the home group (64.8 +/- 14.2, p < 0.05). The home group showed milder leg paresis (p < 0.01), better Barthel index (p < 0.01), a higher number of family members (p < 0.05), and a better substantial care ability (p < 0.01) than LTCF group. 3) Three factors, higher Barthel index (odds ratio: 1.36), higher number of family members (1.84), and better substantial care ability (1.94), were found to facilitate discharge to home. Two other factors, hemorrhage (0.39) and public assistance (0.04), adversely affected the likelihood of discharge to home. CONCLUSIONS: This study suggests that a stroke unit could accelerate discharge home by improving the Barthel index. In addition, socioeconomic factors should be taken into consideration.     

The anti-ischemic effects of CP-060S during pacing-induced ischemia in anesthetized dogs

CP-060 S, (-)-( S)-2-[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl]-3-[3-[N-methyl-N-[2-(3 ,4-methylenedioxyphenoxy)ethyl]-amino]propyl]-1,3-thiazolidin++ +-4-one hydrogen fumarate, is a novel cardioprotective drug which prevents Na+-, Ca2+-overload and has Ca2+ channel blocking activity. We compared the anti-ischemic effects of CP-060S with those of diltiazem, a Ca2+ channel blocker, and R56865, N-[1-[4-(4-fluorophenoxy)butyl]-4-piperidinyl]-N-methyl-2-benzothiazo lamine, a Na+-, Ca2+-overload inhibitor, in a canine pacing-induced ischemia model. CP-060S 100 microg kg(-1) significantly suppressed the pacing-induced ischemic epicardial ST-segment elevation by maximally 75%, while diltiazem 100 microg kg(-1) suppressed it by maximally 35%. R56865 100 microg kg(-1) significantly suppressed the ST-segment elevation by maximally 30%. In addition, diltiazem 100 microg kg(-1) caused synergistic suppression of ST-segment elevation by 70% when administered simultaneously with R56865 100 microg kg(-1). These results suggest that a Na+-, Ca2+-overload preventive action and a Ca2+ channel blocking action independently contribute to the suppression of the ST-segment elevation. Therefore, CP-060S may suppress pacing-induced ST-segment elevation by a dual action by preventing Na+-, Ca2+-overload and the Ca2+ channel blockade.     

Purification and Partial Characterization of an Indomethacin Hydrolyzing Enzyme from Pig Liver

Purpose. Indomethacin is well known to be metabolized via O-demethylation and N-deacylation. In this paper we found an enzyme involved in the hydrolysis of amide-linkage of indomethacin and partially characterized it as well as its substrate specificity. Methods. An indomethacin hydrolyzing enzyme was purified to homogeneity from pig liver microsomes using columns of Q-Sepharose, Red-Sepharose and Blue-Sepharose. The enzyme activity was assayed by measuring of -chlorobenzoic acid liberated from indomethacin by HPLC. Results. The purified enzyme effectively hydrolyzed the amide linkage in indomethacin but not those in -naphthylacetate and -nitrophenylacetate, which are typical substrates for carboxylesterase. The subunit molecular mass of the enzyme was 65 kDa according SDS-polyacrylamide gel electrophoresis. The Michaelis constant (Km) and maximum velocity (Vmax) values for indomethacin were 67.8 µM and 9.02 nmol/min/mg protein, respectively. The amino acid sequence analysis of the enzyme after cyanogen bromide cleavage showed high homology with a mouse carboxylesterase isozyme designated as ES-male. The activity of indomethacin hydrolysis was relatively high in the pig, rabbit and human liver homogenate, but not in those from rat and mouse. On the other hand, purified human liver carboxylesterases pl 5.3 and 4.5, and pig liver carboxylesterases have no catalytic activity for indomethacin. Conclusions. These results indicate that the hydrolysis of amide-linkage of indomethacin in humans would be associated with an enzyme similar to the indomethacin hydrolyzing enzyme from pig liver microsomes described here.     

Progression of cellular repopulation and collagen synthesis in fresh-frozen allograft tendons.

The progression of cellular repopulation and collagen synthesis in fresh-frozen rat patellar tendon allografts was investigated by means of indirect immunofluorescence histologic analysis and electron microscopic techniques for 8 weeks after transplantation. In each of 10 procedures, the medial half of the patellar tendon with a tibial bone block was harvested from a Wistar rat and transplanted into a corresponding defect in the medial patellar tendon of a Lewis rat. Actin filaments in the repopulating cells and newly synthesized collagen fibrils in the graft were identified with rhodamine-phalloidin stain and with a polyclonal antibody against type III collagen aminopropeptide. On the first day after transplantation, no specific fluorescence was detected in the graft. One week later, specific labeling for fibrillar-actin (F-actin) and type III collagen aminopropeptide was detected in an area extending from the adjacent granulation tissue into the proximal end of the graft. F-actin and type III collagen aminopropeptide were aligned along the longitudinal axis of the graft and extended from the proximal suture site toward the distal portion. Two weeks after transplantation, fibrillar labeling for F-actin and type III collagen amino-propeptide showed that remodeling had extended to the midportion of the graft. Labeling throughout the entire graft was detected 4 weeks after transplantation. During the entire remodeling process, the repopulated fibroblasts consistently retained their elongated shape and their alignment with the longitudinal axis of the graft. The cells developed well-organized actin bundles at their peripheries, which identified them as having a myofibroblast phenotype. Immunofluorescence detection for type III collagen aminopropeptide also showed consistent alignment parallel to the longitudinal axis of the graft and a fibrillar arrangement. Electron microscopy revealed thinner collagen fibrils in the vicinity of the fibroblasts, which were aligned in the direction of the actin bundles. These results indicate that, during the early remodeling phase, collagen synthesis and deposition in the graft proceeds while the original alignment of the graft matrix is preserved. The close association between the alignment of actin bundles in repopulated "myofibroblastic" cells and that of newly synthesized collagen fibrils along the lines of the graft matrix may represent evidence of force transmission between the actin cytoskeleton and the linking extracellular matrix in vivo.